Method and reagent for the treatment of asthma and allergic conditions

ABSTRACT

The present invention relates to nucleic acid molecules, including antisense, enzymatic nucleic acid molecules, and RNA interference molecules, such as hammerhead ribozymes, DNAzymes, allozymes, siRNA, decoys and antisense, which modulate the expression of prostaglandin D2 (PTGDS), prostaglandin D2 receptor (PTGDR), and adenosine receptor genes.

PRIORITY

[0001] This application claims the benefit of U.S. Application Ser. No. 60/315,315, filed on Aug. 28, 2001, which is herein incorporated by reference in its entirity.

BACKGROUND OF THE INVENTION

[0002] The present invention relates to therapeutic compositions and methods for the treatment or diagnosis of diseases or conditions related to allergic response. Specifically, the invention provides compositions and methods for the treatment of diseases or conditions related to levels of factors involved in allergic conditions such as asthma, for example prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS) and adenosine A1 receptor (ADORA1). The discussion is provided only for understanding of the invention that follows. This summary is not an admission that any of the work described below is prior art to the claimed invention.

[0003] Asthma is a chronic inflammatory disorder of the lungs characterized by airflow obstruction, bronchial hyper-responsiveness, and airway inflammation. T-lymphocytes that produce TH2 cytokines and cosinophilic leukocytes infiltrate the airways. In the airway and in bronchial alveolar lavage (BAL) fluid of individuals with asthma, high concentrations of TH2 cytokines, interleukin-4 (IL-4), IL-5, and IL-13, are present along with increased levels of adenosine. In contrast to normal individuals, asthmatics respond to adenosine challenge with marked airway obstruction. Upon allergen challenge, mast cells are activated by cross-linked IgE-allergen complexes. Large amounts of prostaglandin D2 (PGD2), the major cyclooxygenase product of arachidonic acid are released. PGD2 is generated from PGH2 via the activity of prostaglandin D2 synthetase (PTGDS). PGD2 receptors and adenosine A1 receptors are present in the lungs and airway along with various other tissues in response to allergic stimuli (Howarth, 1997, Allergy, 52, 12).

[0004] The significance of PGD2 as a mediator of allergic asthma has been established with the development of mice deficient in the PGD2 receptor (DP). DP is a heterotrimeric GTP-binding protein-coupled, rhodopsin-type receptor specific for PGD2 (Hirata et al., 1994, PNAS USA., 91, 11192). These mice fail to develop airway hyperreactivity and have greatly reduced eosinophil infiltration and cytokine accumulation in response to allergens. Upon allergen challenge mice deficient in the prostaglandin D2 (PGD2) receptor (DP) did not develop airway hyperactivity. Cytokine, lymphocyte and eosinophil accumulation in the lungs were greatly reduced (Matsuoka et al., 2000, Science, 287, 2013). The DP −/− mice exhibited no behavioral, anatomic, or histological abnormalities. Primary immune response is not affected by DP disruption.

[0005] Asthma affects more than 100 million people worldwide and more than 17 million Americans (5% of the population). Since 1980 the incidence has more than doubled and deaths have tripled (5,000 deaths in 1995). Annual asthma-related healthcare costs in the US alone were estimated to exceed $14.5 billion in 2000. Current therapies such as inhalant anti-inflammatories and bronchodilators can be used to treat symptoms, however, these therapies do not prevent or cure asthma.

[0006] Sandberg et al., 2001, Prog. Respir. Res., 31, 370-373, describes ribozyme therapy for asthma and COPD.

[0007] Sullivan et al., International U.S. Pat. No. 5,616,488, describes ribozymes targeting interleukin-5 for treatment and diagnosis of asthma and other inflammatory disorders.

[0008] Stinchcomb et al, International PCT Publication No. WO 95/23225, describes ribozymes and methods for inhibiting the expression of disease related genes including genes associated with asthma.

[0009] Nyce, International PCT Publication Nos. WO 00/62736, WO 00/09525, WO 99/13886, WO 98/23294, WO 96/40266 and U.S. Pat. No. 6,025,339 describe specific antisense oligonucleotides targeting certain mRNAs encoding particular adenosine receptors.

SUMMARY OF THE INVENTION

[0010] The invention features novel nucleic acid-based molecules, for example, enzymatic nucleic acid molecules, allozymes, antisense nucleic acids, 2-5A antisense chimeras, triplex forming oligonucleotides, decoy RNA, dsRNA, siRNA, aptamers, and antisense nucleic acids containing RNA cleaving chemical groups, and methods to modulate gene expression, for example, genes encoding prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), and adenosine receptors (AR) such as adenosine receptor A1, A2a, A2b, and A3. In particular, the instant invention features nucleic-acid based molecules and methods to modulate the expression of PTGDR, PTGDS, and adenosine A1 receptor (ADORA1).

[0011] In one embodiment, the invention features one or more nucleic acid-based molecules and methods that independently or in combination modulate the expression of gene(s) encoding prostaglandin D2 receptors (PTGDR), prostaglandin D2 synthetase (PTGDS) and adenosine receptors such as ADORA1. Specifically, the present invention features nucleic acid molecules that modulate the expression of prostaglandin D2 receptor (PTGDR) gene, for example Genbank Accession Nos. U31332 and U31099, prostaglandin D2 synthetase (PTGDS) gene, for example Genbank Accession No. NM_(—)000954, and Adenosine A1 receptor (ADORA1), for example Genbank Accession No. NM_(—)000674.

[0012] The description below of the various aspects and embodiments is provided with reference to the exemplary prostaglandin D2 receptor (PTGDR), prostaglandin D2 synthetase (PTGDS), and adenosine A1 receptor (ADORA1). However, the various aspects and embodiments are also directed to other genes that express prostaglandin proteins and other receptors involved in allergic reactions. Those additional genes can be analyzed for target sites using the methods described for PTGDS, PTGDR, and ADORA1. Thus, the inhibition and the effects of such inhibition of the other genes can be performed as described herein.

[0013] In another embodiment, the invention features an enzymatic nucleic acid molecule comprising a sequence selected from the group consisting of SEQ ID NOs: 228-454, 831-1206, 1438-1668, 1715-2057, and 2247-2666. In yet another embodiment, the invention features an enzymatic nucleic acid molecule comprising at least one binding arm wherein one or more of said binding arms comprises a sequence complementary to a sequence selected from the group consisting of SEQ ID NOs: 1-227, 455-830, 1207-1437, 1669-1714, and 2058-2246.

[0014] In one embodiment, the invention features an antisense nucleic acid molecule comprising a sequence complementary to a sequence selected from the group consisting of SEQ ID NOs: 1-227, 455-830, 1207-1437, 1669-1714, and 2058-2246.

[0015] In another embodiment, an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acids containing RNA cleaving chemical groups of the invention is adapted to treat asthma.

[0016] In one embodiment, an enzymatic nucleic acid molecule of the invention has an endonuclease activity to cleave RNA encoded by a PTGDS and/or PTGDR gene.

[0017] In another embodiment, an enzymatic nucleic acid molecule of the invention is in a hammerhead, Inozyme, Zinzyme, DNAzyme, Amberzyme, or G-cleaver configuration.

[0018] In another embodiment, an enzymatic nucleic acid molecule of the invention having a hammerhead configuration comprises a sequence complementary to a sequence having SEQ ID NOs: 1-227. In yet another embodiment, an enzymatic nucleic acid molecule of invention having a hammerhead configuration comprises a sequence having SEQ ID NOs: 228-454.

[0019] In another embodiment, an enzymatic nucleic acid molecule of the invention having an Inozyme configuration comprises a sequence complementary to a sequence having SEQ ID NOs: 455-830. In yet another embodiment, an enzymatic nucleic acid molecule of invention having an Inozyme configuration comprises a sequence having SEQ ID NOs: 831-1206.

[0020] In another embodiment, an enzymatic nucleic acid molecule of the invention having a Zinzyme configuration comprises a sequence complementary to a sequence having SEQ ID NOs: 1207-1437. In yet another embodiment, an enzymatic nucleic acid molecule of invention having a Zinzyme configuration comprises a sequence having SEQ ID NOs: 1438-1668.

[0021] In another embodiment, an enzymatic nucleic acid molecule of the invention having a DNAzyme configuration comprises a sequence complementary to a sequence having SEQ ID NOs: 1, 13, 55, 69, 74, 104, 112, 120, 123, 128, 131, 138, 147, 154, 157, 158, 169, 188, 192, 208, 221, 463, 475, 489, 505, 527, 541, 552, 554, 561, 563, 572, 591, 601, 605, 627, 637, 645, 652, 653, 661, 668, 669, 670, 676, 692, 699, 706, 719, 725, 732, 737, 741, 747, 763, 774, 782, 800, 805, 807, 816, 818, 823, 827, 828, 1207-1437, and 1669-1714. In yet another embodiment, an enzymatic nucleic acid molecule of invention having a DNAzyme configuration comprises a sequence having SEQ ID NOs: 1715-2057.

[0022] In another embodiment, an enzymatic nucleic acid molecule of the invention having an Amberzyme configuration comprises a sequence complementary to a sequence having SEQ ID NOs: 1207-1437, and 2058-2246. In yet another embodiment, an enzymatic nucleic acid molecule of invention having an Amberzyme configuration comprises a sequence having SEQ ID NOs: 2247-2666.

[0023] In one embodiment, an enzymatic nucleic acid molecule of the invention comprises between 8 and 100 bases complementary to the RNA of PTGDS, ADORA1 and/or PTGDR gene. In another embodiment, an enzymatic nucleic acid molecule of the invention comprises between 14 and 24 bases complementary to a RNA molecule of a PTGDS or PTGDR gene.

[0024] In one embodiment, an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acids containing RNA cleaving chemical groups of the invention is chemically synthesized.

[0025] In another embodiment, an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acids containing RNA cleaving chemical groups of the invention comprises at least one 2′-sugar modification.

[0026] In another embodiment, an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acids containing RNA cleaving chemical groups of the invention comprises at least one nucleic acid base modification.

[0027] In another embodiment, an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acids containing RNA cleaving chemical groups of the invention comprises at least one phosphate backbone modification.

[0028] In one embodiment, the invention features a mammalian cell, for example a human cell, including the enzymatic nucleic acid molecule of the invention.

[0029] In another embodiment, the invention features a method of reducing PTGDS, ADORA1 and/or PTGDR expression or activity in a cell, comprising contacting the cell with an enzymatic nucleic acid molecule of the invention, under conditions suitable for the reduction.

[0030] In another embodiment, the invention features a method of reducing PTGDS, ADORA1 and/or PTGDR expression or activity in a cell, comprising the step of contacting the cell with an antisense nucleic acid molecule of the invention under conditions suitable for the reduction.

[0031] In yet another embodiment, the invention features a method of treatment of a patient having a condition associated with the level of PTGDS, ADORA1 and/or PTGDR, comprising contacting cells of the patient with an enzymatic nucleic acid molecule of the invention, under conditions suitable for the treatment.

[0032] In one embodiment, the invention features a method of treatment of a patient having a condition associated with the level of PTGDS, ADORA1 and/or PTGDR, comprising contacting cells of the patient with an antisense nucleic acid molecule of the invention, under conditions suitable for the treatment.

[0033] In another embodiment, a method of treatment of a patient having a condition associated with the level of PTGDS, ADORA1 and/or PTGDR is featured, wherein the method further comprises the use of one or more drug therapies under conditions suitable for the treatment.

[0034] For example, in one embodiment, the invention features a method for treatment of asthma, allergic rhinitis, or atopic dermatitis under conditions suitable for the treatment.

[0035] In another embodiment, the invention features a method of cleaving a RNA molecule of PTGDS, ADORA1 and/or PTGDR gene comprising contacting an enzymatic nucleic acid molecule of the invention with a RNA molecule of a PTGDS, ADORA1 and/or PTGDR gene under conditions suitable for the cleavage, for example, wherein the cleavage is carried out in the presence of a divalent cation, such as Mg²⁺.

[0036] In one embodiment, an enzymatic nucleic acid molecule of the invention comprises a cap structure, for example a 3′,3′-linked or 5′,5′-linked deoxyabasic ribose derivative, wherein the cap structure is at the 5′-end, or 3′-end, or both the 5′-end and the 3′-end of the enzymatic nucleic acid molecule.

[0037] In another embodiment, an antisense nucleic acid molecule of the invention comprises a cap structure, for example a 3′,3′-linked or 5′,5′-linked deoxyabasic ribose derivative, wherein the cap structure is at the 5′-end, or 3′-end, or both the 5′-end and the 3′-end of the antisense nucleic acid molecule.

[0038] In one embodiment, the invention features an expression vector comprising a nucleic acid sequence encoding at least one enzymatic nucleic acid molecule of the invention, in a manner which allows expression of the nucleic acid molecule.

[0039] In another embodiment, the invention features a mammalian cell, for example, a human cell, including an expression vector of the invention.

[0040] In yet another embodiment, the expression vector of the invention further comprises a sequence for an antisense nucleic acid molecule complementary to a RNA molecule of a PTGDS, ADORA1 and/or PTGDR gene.

[0041] In one embodiment, an expression vector of the invention comprises a nucleic acid sequence encoding two or more enzymatic nucleic acid molecules, which can be the same or different.

[0042] In another embodiment, the invention features a method for treatment of asthma, allergic rhinitis, or atopic dermatitis, comprising administering to a patient an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acid containing RNA cleaving chemical groups of the invention, under conditions suitable for the treatment, including administering to the patient one or more other therapies, for example, inhalant anti-inflammatories, bronchodilators, adenosine inhibitors and adenosine A1 receptor inhibitors.

[0043] In one embodiment, the method of treatment features an enzymatic nucleic acid molecule or antisense nucleic acid molecule of the invention comprises at least five ribose residues, at least ten 2′-O-methyl modifications, and a 3′-end modification, such as a 3′-3′ inverted abasic moiety. In another embodiment, an enzymatic nucleic acid molecule or antisense nucleic acid molecule of the invention further comprises phosphorothioate linkages on at least three of the 5′ terminal nucleotides.

[0044] In another embodiment, the invention features a method of administering to a mammal, for example a human, an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acid containing RNA cleaving chemical groups of the invention, comprising contacting the mammal with the nucleic acid molecule under conditions suitable for the administration, for example, in the presence of a delivery reagent such as a lipid, cationic lipid, phospholipid, or liposome.

[0045] In yet another embodiment, the invention features a method of administering to a mammal an enzymatic nucleic acid molecule, antisense nucleic acid molecule, 2-5A antisense chimera, triplex forming oligonucleotide, decoy RNA, dsRNA, siRNA, aptamer, or antisense nucleic acid containing RNA cleaving chemical groups of the invention in conjunction with a therapeutic agent, comprising contacting the mammal, for example a human, with the nucleic acid molecule and the therapeutic agent under conditions suitable for the administration.

[0046] In one embodiment, the invention features the use of an enzymatic nucleic acid molecule, which can be in a hammerhead, NCH, G-cleaver, Amberzyme, Zinzyme, and/or DNAzyme motif, to down-regulate the expression of a PTGDS, an ADORA1 and/or a PTGDR gene.

[0047] By “inhibit”, “down-regulate”, or “reduce”, it is meant that the expression of the gene, or level of RNA molecules or equivalent RNA molecules encoding one or more proteins or protein subunits, or activity of one or more proteins or protein subunits, such as PTGDS, ADORA1 and/or PTGDR proteins or PTGDS, ADORA1 and/or PTGDR subunit(s), is reduced below that observed in the absence of the nucleic acid molecules of the invention. In one embodiment, inhibition, down-regulation or reduction with an enzymatic nucleic acid molecule is below that level observed in the presence of an enzymatically inactive or attenuated molecule that is able to bind to the same site on the target RNA molecule, but is unable to cleave that RNA molecule. In another embodiment, inhibition, down-regulation, or reduction with antisense oligonucleotides is below that level observed in the presence of, for example, an oligonucleotide with scrambled sequence or with mismatches. In another embodiment, inhibition, down-regulation, or reduction of PTGDS, ADORA1 and/or PTGDR with a nucleic acid molecule of the instant invention is greater in the presence of the nucleic acid molecule than in its absence.

[0048] By “up-regulate” is meant that the expression of a gene, or level of RNA molecules or equivalent RNA molecules encoding one or more proteins, protein subunits, or activity of one or more proteins or protein subunits, such as PTGDS, ADORA1 and/or PTGDR proteins or PTGDS, ADORA1 and/or PTGDR subunits, is greater than that observed in the absence of the nucleic acid molecules of the invention. For example, the expression of a gene, such as PTGDS, ADORA1 and/or PTGDR gene, can be increased in order to treat, prevent, ameliorate, or modulate a pathological condition caused or exacerbated by an absence or low level of gene expression.

[0049] By “modulate” is meant that the expression of the gene, or level of RNA molecules or equivalent RNA molecules encoding one or more protein subunits, or activity of one or more protein subunits is up-regulated or down-regulated, such that the expression, level, or activity is greater than or less than that observed in the absence of a nucleic acid molecule of the invention.

[0050] By “enzymatic nucleic acid molecule” it is meant a nucleic acid molecule that has complementarity in a substrate binding region to a specified gene target, and also has an enzymatic activity that is active to specifically cleave target a RNA molecule. That is, the enzymatic nucleic acid molecule is able to intermolecularly cleave a RNA molecule and thereby inactivate a target RNA molecule. These complementary regions allow sufficient hybridization of an enzymatic nucleic acid molecule to a target RNA molecule and thus permit cleavage. One hundred percent complementarity is preferred, but complementarity as low as 50-75% can also be useful in this invention (see for example Werner and Uhlenbeck, 1995, Nucleic Acids Research, 23, 2092-2096; Hammann et al., 1999, Antisense and Nucleic Acid Drug Dev., 9, 25-31). The nucleic acids can be modified at the base, sugar, and/or phosphate groups. The term enzymatic nucleic acid is used interchangeably with phrases such as ribozymes, catalytic RNA, enzymatic RNA, catalytic DNA, aptazyme or aptamer-binding ribozyme, regulatable ribozyme, catalytic oligonucleotides, nucleozyme, DNAzyme, RNA enzyme, endoribonuclease, endonuclease, minizyme, leadzyme, oligozyme or DNA enzyme. All of these terminologies describe nucleic acid molecules with enzymatic activity. The specific enzymatic nucleic acid molecules described in the instant application are not limiting in the invention and those skilled in the art will recognize that all that is important in an enzymatic nucleic acid molecule of this invention is that it has a specific substrate binding site that is complementary to one or more of the target nucleic acid regions, and that it have nucleotide sequences within or surrounding that substrate binding site which impart a nucleic acid cleaving and/or ligation activity to the molecule (Cech et al, U.S. Pat. No. 4,987,071; Cech et al., 1988, 260 JAMA 3030).

[0051] By “nucleic acid molecule” as used herein is meant a molecule having nucleotides.

[0052] The nucleic acid can be single, double, or multiple stranded and can comprise modified or unmodified nucleotides or non-nucleotides or various mixtures and combinations thereof.

[0053] By “enzymatic portion” or “catalytic domain” is meant that portion/region of the enzymatic nucleic acid molecule essential for cleavage of a nucleic acid substrate (for example see FIGS. 1-4).

[0054] By “substrate binding arm” or “substrate binding domain” is meant that portion/region of a enzymatic nucleic acid that is able to interact, for example via complementarity (i.e., able to base-pair with), with a portion of its substrate. Such complementarity can be 100%, but can be less if desired. For example, as few as 10 bases out of 14 can be base-paired (see for example Werner and Uhlenbeck, 1995, Nucleic Acids Research, 23, 2092-2096; Hammann et al, 1999, Antisense and Nucleic Acid Drug Dev., 9, 25-31). Examples of such arms are shown generally in FIGS. 1-4. That is, these arms contain sequences within a enzymatic nucleic acid that are intended to bring enzymatic nucleic acid and target RNA together through complementary base-pairing interactions. The enzymatic nucleic acid of the invention can have binding arms that are contiguous or non-contiguous and can be of varying lengths. The length of the binding arm(s) can be greater than or equal to four nucleotides and of sufficient length to stably interact with a target RNA; in one embodiment they can be 12-100 nucleotides; in another embodiment they can be 14-24 nucleotides long (see for example Werner and Uhlenbeck, supra; Hamman et al., supra; Hampel et al., EP0360257; Berzal-Herranze et al., 1993, EMBO J., 12, 2567-73) or between 8 and 14 nucleotides long. If two binding arms are chosen, the design is such that the length of the binding arms are symmetrical (i.e., each of the binding arms is of the same length; e.g., four and four, five and five nucleotides, or six and six nucleotides, or seven and seven nucleotides long) or asymmetrical (i.e., the binding arms are of different length; e.g., three and five, six and three nucleotides; three and six nucleotides long; four and five nucleotides long; four and six nucleotides long; four and seven nucleotides long; and the like).

[0055] By “Inozyme” or “NCH” motif or configuration is meant, an enzymatic nucleic acid molecule comprising a motif as is generally described as NCH Rz in FIG. 1. Inozymes possess endonuclease activity to cleave RNA substrates having a cleavage triplet NCH/, where N is a nucleotide, C is cytidine and H is adenosine, uridine or cytidine, and / represents the cleavage site. H is used interchangeably with X. Inozymes can also possess endonuclease activity to cleave RNA substrates having a cleavage triplet NCN/, where N is a nucleotide, C is cytidine, and / represents the cleavage site. “I” in FIG. 1 represents an Inosine nucleotide, including a ribo-Inosine or xylo-Inosine nucleoside.

[0056] By “G-cleaver” motif or configuration is meant, an enzymatic nucleic acid molecule comprising a motif as is generally described as G-cleaver Rz in FIG. 1. G-cleavers possess endonuclease activity to cleave RNA substrates having a cleavage triplet NYN/, where N is a nucleotide, Y is uridine or cytidine and/represents the cleavage site. G-cleavers can be chemically modified as is generally shown in FIG. 1.

[0057] By “amberzyme” motif or configuration is meant, an enzymatic nucleic acid molecule comprising a motif as is generally described in FIG. 2. Amberzymes possess endonuclease activity to cleave RNA substrates having a cleavage triplet NG/N, where N is a nucleotide, G is guanosine, and/represents the cleavage site. Amberzymes can be chemically modified to increase nuclease stability through substitutions as are generally shown in FIG. 2. In addition, differing nucleoside and/or non-nucleoside linkers can be used to substitute the 5′-gaaa-3′ loops shown in the figure. Amberzymes represent a non-limiting example of an enzymatic nucleic acid molecule that does not require a ribonucleotide (2′-OH) group within its own nucleic acid sequence for activity.

[0058] By “zinzyme” motif or configuration is meant, an enzymatic nucleic acid molecule comprising a motif as is generally described in FIG. 3. Zinzymes possess endonuclease activity to cleave RNA substrates having a cleavage triplet including but not limited to YG/Y, where Y is uridine or cytidine, and G is guanosine and/represents the cleavage site.

[0059] Zinzymes can be chemically modified to increase nuclease stability through substitutions as are generally shown in FIG. 3, including substituting 2′-O-methyl guanosine nucleotides for guanosine nucleotides. In addition, differing nucleotide and/or non-nucleotide linkers can be used to substitute the 5′-gaaa-2′ loop shown in the figure. Zinzymes represent a non-limiting example of an enzymatic nucleic acid molecule that does not require a ribonucleotide (2′-OH) group within its own nucleic acid sequence for activity.

[0060] By ‘DNAzyme’ is meant, an enzymatic nucleic acid molecule that does not require the presence of a 2′-OH group within its own nucleic acid sequence for activity. In particular embodiments the enzymatic nucleic acid molecule can have an attached linker or linkers or other attached or associated groups, moieties, or chains containing one or more nucleotides with 2′-OH groups. DNAzymes can be synthesized chemically or expressed endogenously in vivo, by means of a single stranded DNA vector or equivalent thereof. An example of a DNAzyme is shown in FIG. 4 and is generally reviewed in Usman et al., U.S. Pat. No. 6,159,714; Chartrand et al., 1995, NAR 23, 4092; Breaker et al., 1995, Chem. Bio. 2, 655; Santoro et al., 1997, PNAS 94, 4262; Breaker, 1999, Nature Biotechnology, 17, 422-423; and Santoro et. al., 2000, J. Am. Chem. Soc., 122, 2433-39. The “10-23” DNAzyme motif is one particular type of DNAzyme that was evolved using in vitro selection (see Santoro et al., supra). Additional DNAzyme motifs can be selected for using techniques similar to those described in these references, and hence, are within the scope of the present invention.

[0061] By “sufficient length” is meant an oligonucleotide of greater than or equal to 3 nucleotides that is of a length great enough to provide the intended function under the expected condition. For example, for binding arms of enzymatic nucleic acid “sufficient length” means that the binding arm sequence is long enough to provide stable binding to a target site under the expected binding conditions. The binding arms are not so long as to prevent useful turnover of the nucleic acid molecule.

[0062] By “stably interact” is meant interaction of the oligonucleotides with target nucleic acid (e.g., by forming hydrogen bonds with complementary nucleotides in the target under physiological conditions) that is sufficient to the intended purpose (e.g., cleavage of target RNA by an enzyme).

[0063] By “equivalent” RNA to PTGDS is meant to include RNA molecules having homology (partial or complete) to RNA molecules encoding PTGDS proteins or encoding proteins with similar function as PTGDS proteins in various organisms, including human, rodent, primate, rabbit, pig, plants, protozoans, fungi, and other microorganisms and parasites. The equivalent RNA sequence can also include in addition to the coding region, regions such as 5′-untranslated region, 3′-untranslated region, introns, intron-exon junction and the like.

[0064] By “equivalent” RNA to PTGDR is meant to include RNA molecules having homology (partial or complete) to RNA molecules encoding PTGDR proteins or encoding proteins with similar function as PTGDR proteins in various organisms, including human, rodent, primate, rabbit, pig, plants, protozoans, fungi, and other microorganisms and parasites. The equivalent RNA sequence can also include in addition to the coding region, regions such as 5′-untranslated region, 3′-untranslated region, introns, intron-exon junction and the like.

[0065] By “equivalent” RNA to ADORA1 is meant to include RNA molecules having homology (partial or complete) to RNA molecule encoding ADORA1 proteins or encoding proteins with similar function as ADORA1 proteins in various organisms, including human, rodent, primate, rabbit, pig, plants, protozoans, fungi, and other microorganisms and parasites. The equivalent RNA sequence can also include in addition to the coding region, regions such as 5′-untranslated region, 3′-untranslated region, introns, intron-exon junction and the like.

[0066] By “homology” is meant the nucleotide sequence of two or more nucleic acid molecules is partially or completely identical.

[0067] By “antisense nucleic acid”, it is meant a non-enzymatic nucleic acid molecule that binds to target RNA by means of RNA-RNA or RNA-DNA or RNA-PNA (protein nucleic acid; Egholm et al., 1993 Nature 365, 566) interactions and alters the activity of the target RNA (for a review, see Stein and Cheng, 1993 Science 261, 1004 and Woolf et al., U.S. Pat. No. 5,849,902). Typically, antisense molecules are complementary to a target sequence along a single contiguous sequence of the antisense molecule. However, in certain embodiments, an antisense molecule can bind to substrate such that the substrate molecule forms a loop, and/or an antisense molecule can bind such that the antisense molecule forms a loop. Thus, the antisense molecule can be complementary to two (or even more) non-contiguous substrate sequences or two (or even more) non-contiguous sequence portions of an antisense molecule can be complementary to a target sequence or both. For a review of current antisense strategies, see Schmajuk et al., 1999, J. Biol. Chem., 274, 21783-21789, Delihas et al., 1997, Nature, 15, 751-753, Stein et al., 1997, Antisense N. A. Drug Dev., 7, 151, Crooke, 2000, Methods Enzymol., 313, 3-45; Crooke, 1998, Biotech. Genet. Eng. Rev., 15, 121-157, Crooke, 1997, Ad. Pharmacol., 40, 1-49. In addition, antisense DNA can be used to target RNA by means of DNA-RNA interactions, thereby activating RNase H, which digests the target RNA in the duplex. The antisense oligonucleotides can comprise one or more RNAse H activating region, which is capable of activating RNAse H cleavage of a target RNA. Antisense DNA can be synthesized chemically or expressed via the use of a single stranded DNA expression vector or equivalent thereof.

[0068] By “RNase H activating region” is meant a region (generally greater than or equal to 4-25 nucleotides in length, and in one embodiment from 5-11 nucleotides in length) of a nucleic acid molecule capable of binding to a target RNA to form a non-covalent complex that is recognized by cellular RNase H enzyme (see for example Arrow et al., U.S. Pat. No. 5,849,902; Arrow et al., U.S. Pat. No. 5,989,912). The RNase H enzyme binds to the nucleic acid molecule-target RNA complex and cleaves the target RNA sequence. The RNase H activating region comprises, for example, phosphodiester, phosphorothioate (at least four of the nucleotides are phosphorothiote substitutions; and in another embodiment, 4-11 of the nucleotides are phosphorothiote substitutions); phosphorodithioate, 5′-thiophosphate, or methylphosphonate backbone chemistry or a combination thereof. In addition to one or more backbone chemistries described above, the RNase H activating region can also comprise a variety of sugar chemistries. For example, the RNase H activating region can comprise deoxyribose, arabino, fluoroarabino or a combination thereof, nucleotide sugar chemistry. Those skilled in the art will recognize that the foregoing are non-limiting examples and that any combination of phosphate, sugar and base chemistry of a nucleic acid that supports the activity of RNase H enzyme is within the scope of the definition of the RNase H activating region and the instant invention.

[0069] By “2-5A antisense chimera” is meant an antisense oligonucleotide containing a 5′-phosphorylated 2′-5′-linked adenylate residue. These chimeras bind to target RNA in a sequence-specific manner and activate a cellular 2-5A-dependent ribonuclease which, in turn, cleaves the target RNA (Torrence et al., 1993 Proc. Natl. Acad. Sci. USA 90, 1300; Silverman et al., 2000, Methods Enzymol., 313, 522-533; Player and Torrence, 1998, Pharmacol. Ther., 78, 55-113).

[0070] By “triplex forming oligonucleotides” is meant an oligonucleotide that can bind to a double-stranded DNA in a sequence-specific manner to form a triple-strand helix. Formation of such triple helix structure has been shown to inhibit transcription of the targeted gene (Duval-Valentin et al., 1992 Proc. Natl. Acad. Sci. USA 89, 504; Fox, 2000, Curr. Med. Chem., 7, 17-37; Praseuth et. al., 2000, Biochim. Biophys. Acta, 1489, 181-206).

[0071] By “gene” it is meant a nucleic acid that encodes an RNA, for example, nucleic acid sequences including but not limited to structural genes encoding a polypeptide.

[0072] “Complementarity” refers to the ability of a nucleic acid to form hydrogen bond(s) with another RNA molecule by either traditional Watson-Crick or other non-traditional types. In reference to the nucleic molecules of the present invention, the binding free energy for a nucleic acid molecule with its target or complementary sequence is sufficient to allow the relevant function of the nucleic acid to proceed, e.g., enzymatic nucleic acid cleavage, antisense or triple helix inhibition. Determination of binding free energies for nucleic acid molecules is well known in the art (see, e.g., Turner et al., 1987, CSHSymp. Quant. Biol. LII pp.123-133; Frier et al., 1986, Proc. Nat. Acad. Sci. USA 83:9373-9377; Turner et al., 1987, J. Am. Chem. Soc. 109:3783-3785). A percent complementarity indicates the percentage of contiguous residues in a nucleic acid molecule that can form hydrogen bonds (e.g., Watson-Crick base pairing) with a second nucleic acid sequence (e.g., 5, 6, 7, 8, 9, 10 out of 10 being 50%, 60%, 70%, 80%, 90%, and 100% complementary). “Perfectly complementary” means that all the contiguous residues of a nucleic acid sequence will hydrogen bond with the same number of contiguous residues in a second nucleic acid sequence.

[0073] By “RNA” is meant a molecule comprising at least one ribonucleotide residue. By “ribonucleotide” or “2′-OH” is meant a nucleotide with a hydroxyl group at the 2′ position of a β-D-ribo-furanose moiety.

[0074] By “decoy RNA” is meant an RNA molecule or aptamer that is designed to preferentially bind to a predetermined ligand. Such binding can result in the inhibition or activation of a target molecule. The decoy RNA or aptamer can compete with a naturally occurring binding target for the binding of a specific ligand. For example, it has been shown that over-expression of HIV trans-activation response (TAR) RNA can act as a “decoy” and efficiently binds HIV tat protein, thereby preventing it from binding to TAR sequences encoded in the HIV RNA (Sullenger et al., 1990, Cell, 63, 601-608). This is but a specific example and those in the art will recognize that other embodiments can be readily generated using techniques generally known in the art, see for example Gold et al., 1995, Annu. Rev. Biochem., 64, 763; Brody and Gold, 2000, J. Biotechnol., 74, 5; Sun, 2000, Curr. Opin. Mol. Ther., 2, 100; Kusser, 2000, J. Biotechnol., 74, 27; Hermann and Patel, 2000, Science, 287, 820; and Jayasena, 1999, Clinical Chemistry, 45, 1628. Similarly, a decoy RNA can be designed to bind to a D2 receptor and block the binding of PTGDS or a decoy RNA can be designed to bind to PTGDS and prevent interaction with the D2 receptor.

[0075] The term “double stranded RNA” or “dsRNA” as used herein refers to a double stranded RNA molecule capable of RNA interference, including short interfering RNA “siRNA” (see, e.g., Bass, 2001, Nature, 411, 428-429; Elbashir et al., 2001, Nature, 411, 494-498).

[0076] The term “allozyme” as used herein refers to an allosteric enzymatic nucleic acid molecule, see, e.g., George et al., U.S. Pat. Nos. 5,834,186 and 5,741,679, Shih et al., U.S. Pat. No. 5,589,332, Nathan et al., U.S. Pat. No. 5,871,914, Nathan and Ellington, International PCT publication No. WO 00/24931, Breaker et al., International PCT Publication Nos. WO 00/26226 and 98/27104, and Sullenger et al., International PCT publication No. WO 99/29842. The term “2-5A chimera” as used herein refers to an oligonucleotide containing a 5′-phosphorylated 2′-5′-linked adenylate residue. These chimeras bind to target RNA in a sequence-specific manner and activate a cellular 2-5A-dependent ribonuclease which, in turn, cleaves the target RNA (Torrence et al., 1993 Proc. Natl. Acad. Sci. USA 90, 1300; Silverman et al., 2000, Methods Enzymol., 313, 522-533; Player and Torrence, 1998, Pharmacol. Ther., 78, 55-113).

[0077] The term “triplex forming oligonucleotides” as used herein refers to an oligonucleotide that can bind to a double-stranded DNA in a sequence-specific manner to form a triple-strand helix. Formation of such triple helix structure has been shown to inhibit transcription of the targeted gene (Duval-Valentin et al., 1992 Proc. Natl. Acad. Sci. USA 89, 504; Fox, 2000, Curr. Med. Chem., 7, 17-37; Praseuth et. al., 2000, Biochim. Biophys. Acta, 1489, 181-206).

[0078] Several varieties of naturally-occurring enzymatic RNAs are known presently. Each can catalyze the hydrolysis of RNA phosphodiester bonds in trans (and thus can cleave other RNA molecules) under physiological conditions. Table I summarizes some of the characteristics of these ribozymes. In general, enzymatic nucleic acids act by first binding to a target RNA. Such binding occurs through the target binding portion of a enzymatic nucleic acid that is held in close proximity to an enzymatic portion of the molecule that acts to cleave the target RNA. Thus, the enzymatic nucleic acid first recognizes and then binds a target RNA through complementary base-pairing, and once bound to the correct site, acts enzymatically to cut the target RNA. Strategic cleavage of such a target RNA will destroy its ability to direct synthesis of an encoded protein. After an enzymatic nucleic acid has bound and cleaved its RNA target, it is released from that RNA to search for another target and can repeatedly bind and cleave new targets. Thus, a single ribozyme molecule is able to cleave many molecules of target RNA. In addition, the ribozyme is a highly specific inhibitor of gene expression, with the specificity of inhibition depending not only on the base-pairing mechanism of binding to the target RNA, but also on the mechanism of target RNA cleavage. Single mismatches, or base-substitutions, near the site of cleavage can completely eliminate catalytic activity of a ribozyme.

[0079] The enzymatic nucleic acid molecule that cleave the specified sites in PTGDS, ADORA1 and PTGDR-specific RNAs represent a novel therapeutic approach to treat a variety of allergic diseases or conditions, including but not limited to asthma, allergic rhinitis, atopic dermatitis, and/or other allergic or inflammatory diseases and conditions which respond to the modulation of PTGDS, ADORA1 and/or PTGDR expression.

[0080] In one embodiment of the inventions described herein, the enzymatic nucleic acid molecule is formed in a hammerhead or hairpin motif, but can also be formed in the motif of a hepatitis delta virus, group I intron, group II intron or RNase P RNA (in association with an RNA guide sequence), Neurospora VS RNA, DNAzymes, NCH cleaving motifs, or G-cleavers. Examples of such hammerhead motifs are described by Dreyfus, supra, Rossi et al., 1992, AIDS Research and Human Retroviruses 8, 183; of hairpin motifs by Hampel et al., EP0360257, Hampel and Tritz, 1989 Biochemistry 28, 4929, Feldstein et al., 1989, Gene 82, 53, Haseloff and Gerlach, 1989, Gene, 82, 43, and Hampel et al., 1990 Nucleic Acids Res. 18, 299; Chowrira & McSwiggen, U.S. Pat. No. 5,631,359; of the hepatitis delta virus motif is described by Perrotta and Been, 1992 Biochemistry 31, 16; of the RNase P motif by Guerrier-Takada et al., 1983 Cell 35, 849; Forster and Altman, 1990, Science 249, 783; Li and Altman, 1996, Nucleic Acids Res. 24, 835; Neurospora VS RNA ribozyme motif is described by Collins (Saville and Collins, 1990 Cell 61, 685-696; Saville and Collins, 1991 Proc. Natl. Acad. Sci. USA 88, 8826-8830; Collins and Olive, 1993 Biochemistry 32, 2795-2799; Guo and Collins, 1995, EMBO. J 14, 363); Group II introns are described by Griffin et al., 1995, Chem. Biol. 2, 761; Michels and Pyle, 1995, Biochemistry 34, 2965; Pyle et al., International PCT Publication No. WO 96/22689; of the Group I intron by Cech et al., U.S. Pat. No. 4,987,071 and of DNAzymes by Usman et al., International PCT Publication No. WO 95/11304; Chartrand et al., 1995, NAR 23, 4092; Breaker et al., 1995, Chem. Bio. 2, 655; Santoro et al., 1997, PNAS 94, 4262, and Beigelman et al., International PCT publication No. WO 99/55857. NCH cleaving motifs are described in Ludwig & Sproat, International PCT Publication No. WO 98/58058; and G-cleavers are described in Kore et al., 1998, Nucleic Acids Research 26, 4116-4120 and Eckstein et al., International PCT Publication No. WO 99/16871. Additional motifs such as the Aptazyme (Breaker et al., WO 98/43993), Amberzyme (Class I motif; FIG. 2; Beigelman et al., U.S. Ser. No. 09/301,511) and Zinzyme (FIG. 3) (Beigelman et al., U.S. Ser. No. 09/301,511), all included by reference herein including drawings, can also be used in the present invention. These specific motifs or configurations are not limiting in the invention and those skilled in the art will recognize that all that is important in an enzymatic nucleic acid molecule of this invention is that it has a specific substrate binding site which is complementary to one or more of the target gene RNA regions, and that it have nucleotide sequences within or surrounding that substrate binding site which impart an RNA cleaving activity to the molecule (Cech et al., U.S. Pat. No. 4,987,071).

[0081] In one embodiment of the present invention, a nucleic acid molecule of the instant invention can be between 12 and 100 nucleotides in length. Exemplary enzymatic nucleic acid molecules of the invention are shown in Table III-VII. For example, enzymatic nucleic acid molecules of the invention can be between 15 and 50 nucleotides in length, and in another embodiment between 25 and 40 nucleotides in length, e.g., 34, 36, or 38 nucleotides in length (for example see Jarvis et al., 1996, J. Biol. Chem., 271, 29107-29112). Exemplary DNAzymes of the invention are can between 15 and 40 nucleotides in length, and in one embodiment, between 25 and 35 nucleotides in length, e.g., 29, 30, 31, or 32 nucleotides in length (see, e.g., Santoro et al., 1998, Biochemistry, 37, 13330-13342; Chartrand et al., 1995, Nucleic Acids Research, 23, 4092-4096). Exemplary antisense molecules of the invention can be between 15 and 75 nucleotides in length, and in one embodiment between 20 and 35 nucleotides in length, e.g., 25, 26, 27, or 28 nucleotides in length (see for example Woolf et al., 1992, PNAS., 89, 7305-7309; Milner et al., 1997, Nature Biotechnology, 15, 537-541). Exemplary triplex forming oligonucleotide molecules of the invention are between 10 and 40 nucleotides in length, and in one embodiment are between 12 and 25 nucleotides in length, e.g., 18, 19, 20, or 21 nucleotides in length (see for example Maher et al., 1990, Biochemistry, 29, 8820-8826; Strobel and Dervan, 1990, Science, 249, 73-75). Those skilled in the art will recognize that all that is required is for the nucleic acid molecule to be of length and conformation sufficient and suitable for the nucleic acid molecule to catalyze a reaction contemplated herein. The length of the nucleic acid molecules of the instant invention are not limiting within the general limits stated.

[0082] In one embodiment, a nucleic acid molecule that modulates, for example, down-regulates, PTGDS replication or expression comprises between 8 and 100 bases complementary to a RNA molecule of PTGDS. In another embodiment, a nucleic acid molecule that modulates PTGDS replication or expression comprises between 14 and 24 bases complementary to a RNA molecule of PTGDS.

[0083] In another embodiment, a nucleic acid molecule that modulates, for example, down-regulates, PTGDR replication or expression comprises between 8 and 100 bases complementary to a RNA molecule of PTGDR. In another embodiment, a nucleic acid molecule that modulates PTGDR replication or expression comprises between 14 and 24 bases complementary to a RNA molecule of PTGDR.

[0084] In another embodiment, a nucleic acid molecule that modulates, for example, down-regulates, ADORA1 replication or expression comprises between 8 and 100 bases complementary to a RNA molecule of ADORA1. In another embodiment, a nucleic acid molecule that modulates ADORA1 replication or expression comprises between 14 and 24 bases complementary to a RNA molecule of ADORA1.

[0085] The invention provides a method for producing a class of nucleic acid-based gene modulating agents that exhibit a high degree of specificity for the RNA of a desired target. For example, the enzymatic nucleic acid molecule is can be targeted to a highly conserved sequence region of target RNAs encoding PTGDS, ADORA1 and/or PTGDR (e.g., PTGDS, ADORA1 and/or PTGDR genes) such that specific treatment of a disease or condition can be provided with either one or several nucleic acid molecules of the invention. Such nucleic acid molecules can be delivered exogenously to specific tissue or cellular targets as required. Alternatively, the nucleic acid molecules (e.g., ribozymes and antisense) can be expressed from DNA and/or RNA vectors that are delivered to specific cells.

[0086] As used in herein “cell” is used in its usual biological sense, and does not refer to an entire multicellular organism. The cell can, for example, be in vitro, e.g., in cell culture, or present in a multicellular organism, including, e.g., birds, plants and mammals such as humans, cows, sheep, apes, monkeys, swine, dogs, and cats. The cell may be prokaryotic (e.g., bacterial cell) or eukaryotic (e.g., mammalian or plant cell).

[0087] By “PTGDR proteins” is meant, a protein receptor or a mutant protein or peptide derivative thereof, having prostaglandin D2 receptor activity, for example, having the ability to bind prostaglandin D2 and/or having GTP-binding protein coupled activity.

[0088] By “PTGDS proteins” is meant, a prostaglandin synthetase protein or a mutant protein or peptide derivative thereof, having prostaglandin D2 synthetase activity, for example, having the ability to convert PGH2 to PGD2.

[0089] By “highly conserved sequence region” is meant, a nucleotide sequence of one or more regions in a target gene does not vary significantly from one generation to the other or from one biological system to the other.

[0090] Nucleic acid-based inhibitors of PTGDS, ADORA1 and PTGDR expression are useful for the prevention and/or treatment of allergic diseases or conditions, including but not limited to asthma, allergic rhinitis, atopic dermatitis, and any other diseases or conditions that are related to or will respond to the levels of PTGDS, ADORA1 and/or PTGDR in a cell or tissue, alone or in combination with other therapies. The reduction of PTGDS, ADORA1 and/or PTGDR expression (specifically PTGDS, ADORA1 and/or PTGDR gene RNA levels) and thus reduction in the level of the respective protein relieves, to some extent, the symptoms of the disease or condition.

[0091] The nucleic acid-based inhibitors of the invention can be added directly, or can be complexed with cationic lipids, packaged within liposomes, or otherwise delivered to target cells or tissues, for example by pulmonary delivery of an aerosol formulation with an inhaler or nebulizer. The nucleic acid or nucleic acid complexes can be locally administered to relevant tissues ex vivo, or in vivo through inhalation, injection or infusion pump, with or without their incorporation in biopolymers. In preferred embodiments, the enzymatic nucleic acid inhibitors comprise sequences that are complementary to the substrate sequences in Tables III to VII. Examples of such enzymatic nucleic acid molecules also are shown in Tables III to VII. Examples of such enzymatic nucleic acid molecules consist essentially of sequences defined in these tables.

[0092] In another embodiment, the invention features antisense nucleic acid molecules and 2-5A chimera including sequences complementary to the substrate sequences shown in Tables III to VII. Such nucleic acid molecules can include sequences as shown for the binding arms of the enzymatic nucleic acid molecules in Tables III to VII. Similarly, triplex molecules can be provided targeted to the corresponding DNA target regions, and containing the DNA equivalent of a target sequence or a sequence complementary to the specified target (substrate) sequence. Typically, antisense molecules are complementary to a target sequence along a single contiguous sequence of the antisense molecule. However, in certain embodiments, an antisense molecule can bind to substrate such that the substrate molecule forms a loop, and/or an antisense molecule can bind such that the antisense molecule forms a loop. Thus, the antisense molecule can be complementary to two (or even more) non-contiguous substrate sequences or two (or even more) non-contiguous sequence portions of an antisense molecule can be complementary to a target sequence or both.

[0093] By “consists essentially of” is meant that the active nucleic acid molecule of the invention, for example, an enzymatic nucleic acid molecule, contains an enzymatic center or core equivalent to those in the examples, and binding arms able to bind RNA such that cleavage at the target site occurs. Other sequences can be present that do not interfere with such cleavage. Thus, a core region can, for example, include one or more loop, stem-loop structure, or linker which does not prevent enzymatic activity. Thus, the underlined regions in the sequences in Tables III and IV can be such a loop, stem-loop, nucleotide linker, and/or non-nucleotide linker and can be represented generally as sequence “X”. For example, a core sequence for a hammerhead enzymatic nucleic acid can comprise a conserved sequence, such as 5′-CUGAUGAG-3′ and 5′-CGAA-3′ connected by “X”, where X is 5′-GCCGUUAGGC-3′ (SEQ ID NO: 2678), or any other Stem II region known in the art, or a nucleotide and/or non-nucleotide linker. Similarly, for other nucleic acid molecules of the instant invention, such as Inozyme, G-cleaver, amberzyme, zinzyme, DNAzyme, antisense, 2-5A antisense, triplex forming nucleic acid, and decoy nucleic acids, other sequences or non-nucleotide linkers can be present that do not interfere with the function of the nucleic acid molecule.

[0094] Sequence X can be a linker of ≧2 nucleotides in length, including 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 26, 30, where the nucleotides can be internally base-paired to form a stem of ≧2 base pairs. Alternatively or in addition, sequence X can be a non-nucleotide linker. In yet another embodiment, the nucleotide linker X can be a nucleic acid aptamer, such as an ATP aptamer, HIV Rev aptamer (RRE), HIV Tat aptamer (TAR) and others (for a review see Gold et al., 1995, Annu. Rev. Biochem., 64, 763; and Szostak & Ellington, 1993, in The RNA World, ed. Gesteland and Atkins, pp. 511, CSH Laboratory Press). A “nucleic acid aptamer” as used herein is meant to indicate a nucleic acid sequence capable of interacting with a ligand. The ligand can be any natural or a synthetic molecule, including but not limited to a resin, metabolites, nucleosides, nucleotides, drugs, toxins, transition state analogs, peptides, lipids, proteins, amino acids, nucleic acid molecules, hormones, carbohydrates, receptors, cells, viruses, bacteria and others.

[0095] In yet another embodiment, the non-nucleotide linker X is as defined herein. The term “non-nucleotide” as used herein include either abasic nucleotide, polyether, polyamine, polyamide, peptide, carbohydrate, lipid, or polyhydrocarbon compounds. Specific examples include those described by Seela and Kaiser, Nucleic Acids Res. 1990, 18:6353 and Nucleic Acids Res. 1987, 15:3113; Cload and Schepartz, J. Am. Chem. Soc. 1991, 113:6324; Richardson and Schepartz, J. Am. Chem. Soc. 1991, 113:5109; Ma et al., Nucleic Acids Res. 1993, 21:2585 and Biochemistry 1993, 32:1751; Durand et al., Nucleic Acids Res. 1990, 18:6353; McCurdy et al., Nucleosides & Nucleotides 1991, 10:287; Jschke et al., Tetrahedron Lett. 1993, 34:301; Ono et al., Biochemistry 1991, 30:9914; Arnold et al., International Publication No. WO 89/02439; Usman et al., International Publication No. WO 95/06731; Dudycz et al., International Publication No. WO 95/11910 and Ferentz and Verdine, J. Am. Chem. Soc. 1991, 113:4000, all hereby incorporated by reference herein. A “non-nucleotide” further means any group or compound that can be incorporated into a nucleic acid chain in the place of one or more nucleotide units, including either sugar and/or phosphate substitutions, and allows the remaining bases to exhibit their enzymatic activity. The group or compound can be abasic in that it does not contain a commonly recognized nucleotide base, such as adenosine, guanine, cytosine, uracil or thymine. Thus, in a preferred embodiment, the invention features an enzymatic nucleic acid molecule having one or more non-nucleotide moieties, and having enzymatic activity to cleave an RNA or DNA molecule.

[0096] In another aspect of the invention, enzymatic nucleic acid molecules or antisense molecules that interact with target RNA molecules and down-regulate PTGDS, ADORA1 and/or PTGDR (e.g., PTGDS, ADORA1 and/or PTGDR gene) activity are expressed from transcription units inserted into DNA or RNA vectors. The recombinant vectors can be DNA plasmids or viral vectors. Enzymatic nucleic acid molecule or antisense expressing viral vectors can be constructed based on, but not limited to, adeno-associated virus, retrovirus, adenovirus, or alphavirus. The recombinant vectors capable of expressing the enzymatic nucleic acid molecules or antisense can be delivered as described above, and persist in target cells. Alternatively, viral vectors can be used that provide for transient expression of enzymatic nucleic acid molecules or antisense. Such vectors can be repeatedly administered as necessary. Once expressed, the enzymatic nucleic acid molecules or antisense bind to the target RNA and down-regulate its function or expression. Delivery of enzymatic nucleic acid molecule or antisense expressing vectors can be systemic, such as by intravenous or intramuscular administration, by administration to target cells ex-planted from the patient followed by reintroduction into the patient, or by any other means that would allow for introduction into the desired target cell. Antisense DNA can be expressed via the use of a single stranded DNA intracellular expression vector.

[0097] By “vectors” is meant any nucleic acid- and/or viral-based technique used to deliver a desired nucleic acid.

[0098] By “patient” is meant an organism, which is a donor or recipient of explanted cells, or the cells themselves. “Patient” also refers to an organism to which the nucleic acid molecules of the invention can be administered. A patient can be a mammal or mammalian cells. In one embodiment, a patient is a human or human cells.

[0099] By “enhanced enzymatic activity” is meant to include activity measured in cells and/or in vivo where the activity is a reflection of both the catalytic activity and the stability of the nucleic acid molecules of the invention. In this invention, the product of these properties can be increased in vivo compared to an all RNA enzymatic nucleic acid or all DNA enzyme. In some cases, the activity or stability of the nucleic acid molecule can be decreased (i.e., less than ten-fold), but the overall activity of the nucleic acid molecule is enhanced, in vivo.

[0100] The nucleic acid molecules of the instant invention, individually, or in combination or in conjunction with other drugs, can be used to treat diseases or conditions discussed above. For example, to treat a disease or condition associated with the levels of PTGDS, ADORA1 and/or PTGDR, the patient can be treated, or other appropriate cells can be treated, as is evident to those skilled in the art, individually or in combination with one or more drugs under conditions suitable for the treatment.

[0101] In a further embodiment, the described molecules, such as antisense or enzymatic nucleic acid molecules, can be used in combination with other known treatments to treat conditions or diseases discussed above. For example, the described molecules can be used in combination with one or more known therapeutic agents to treat allergic diseases or conditions, including but not limited to asthma, allergic rhinitis, atopic dermatitis, and/or other allergic or inflammatory diseases and conditions which respond to the modulation of PTGDS, ADORA1 and/or PTGDR expression.

[0102] In another embodiment, the invention features nucleic acid-based inhibitors (e.g., enzymatic nucleic acid molecules (e.g., ribozymes), antisense nucleic acids, 2-5A antisense chimeras, triplex DNA, antisense nucleic acids containing RNA cleaving chemical groups) and methods for their use to down regulate or inhibit the expression of genes (e.g., PTGDS, ADORA1 and/or PTGDR) capable of progression and/or maintenance allergic diseases or conditions, including but not limited to asthma, allergic rhinitis, atopic dermatitis, and/or other allergic or inflammatory diseases and conditions which respond to the modulation of PTGDS, ADORA1 and/or PTGDR expression.

[0103] By “comprising” is meant including, but not limited to, whatever follows the word “comprising”. Thus, use of the term “comprising” indicates that the listed elements are required or mandatory, but that other elements are optional and may or may not be present. By “consisting of” is meant including, and limited to, whatever follows the phrase “consisting of”. Thus, the phrase “consisting of” indicates that the listed elements are required or mandatory, and that no other elements may be present.

[0104] Other features and advantages of the invention will be apparent from the following description of the preferred embodiments thereof, and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

[0105]FIG. 1 shows examples of chemically stabilized ribozyme motifs. HH Rz, represents hammerhead ribozyme motif (Usman et al., 1996, Curr. Op. Struct. Bio., 1, 527); NCH Rz represents the NCH ribozyme motif (Ludwig & Sproat, International PCT Publication No. WO 98/58058); G-Cleaver, represents G-cleaver ribozyme motif (Kore et al., 1998, Nucleic Acids Research 26, 4116-4120, Eckstein et al., International PCT publication No. WO 99/16871). N or n, represent independently a nucleotide that can be same or different and have complementarity to each other; rI, represents ribo-Inosine nucleotide; arrow indicates the site of cleavage within the target. Position 4 of the HH Rz and the NCH Rz is shown as having 2′-C-allyl modification, but those skilled in the art will recognize that this position can be modified with other modifications well known in the art, so long as such modifications do not significantly inhibit the activity of the ribozyme.

[0106]FIG. 2 shows an example of the Amberzyme ribozyme motif that is chemically stabilized (see for example Beigelman et al., International PCT publication No. WO 99/55857).

[0107]FIG. 3 shows an example of the Zinzyme A ribozyme motif that is chemically stabilized (see for example Beigelman et al., Beigelman et al., International PCT publication No. WO 99/55857).

[0108]FIG. 4 shows an example of a specific DNAzyme motif, commonly referred to as the “10-23 motif”, as described by Santoro et al., 1997, PNAS, 94, 4262.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0109] Nucleic Acid Molecules and Mechanism of Action

[0110] Antisense: Antisense molecules can be modified or unmodified RNA, DNA, or mixed polymer oligonucleotides and primarily function by specifically binding to matching sequences resulting in inhibition of peptide synthesis (Wu-Pong, Nov 1994, BioPharm, 20-33). The antisense oligonucleotide binds to target RNA by Watson Crick base-pairing and blocks gene expression by preventing ribosomal translation of the bound sequences either by steric blocking or by activating RNase H enzyme. Antisense molecules can also alter protein synthesis by interfering with RNA processing or transport from the nucleus into the cytoplasm (Mukhopadhyay & Roth, 1996, Crit. Rev. in Oncogenesis 7, 151-190).

[0111] In addition, binding of single stranded DNA to RNA can result in nuclease degradation of the heteroduplex (Wu-Pong, supra; Crooke, supra). To date, the only backbone modified DNA chemistry which act as substrates for RNase H are phosphorothioates, phosphorodithioates, and borontrifluoridates. Recently it has been reported that 2′-arabino and 2′-fluoro arabino-containing oligos can also activate RNase H activity.

[0112] A number of antisense molecules have been described that utilize novel configurations of chemically modified nucleotides, secondary structure, and/or RNase H substrate domains (Woolf et al., International PCT Publication No. WO 98/13526; Thompson et al., International PCT Publication No. WO 99/54459; Hartmann et al., U.S. S No. 60/101,174, filed on Sep. 21, 1998) all of these are incorporated by reference herein in their entirety.

[0113] In addition, antisense deoxyoligoribonucleotides can be used to target RNA by means of DNA-RNA interactions, thereby activating RNase H, which digests the target RNA in the duplex. Antisense DNA can be expressed via the use of a single stranded DNA intracellular expression vector or equivalents and variations thereof.

[0114] Triplex Forming Oligonucleotides (TFO): Single stranded DNA can be designed to bind to genomic DNA in a sequence specific manner. TFOs are comprised of pyrimidine-rich oligonucleotides which bind DNA helices through Hoogsteen Base-pairing (Wu-Pong, supra). The resulting triple helix composed of the DNA sense, DNA antisense, and TFO disrupts RNA synthesis by RNA polymerase. The TFO mechanism can result in gene expression or cell death since binding can be irreversible (Mukhopadhyay & Roth, supra).

[0115] 2-5A Antisense Chimera: The 2-5A system is an interferon mediated mechanism for RNA degradation found in higher vertebrates (Mitra et al., 1996, Proc Nat Acad Sci USA 93, 6780-6785). Two types of enzymes, 2-5A synthetase and RNase L, are required for RNA cleavage. The 2-5A synthetases require double stranded RNA to form 2′-5′ oligoadenylates (2-5A). 2-5A then acts as an allosteric effector for utilizing RNase L, which has the ability to cleave single stranded RNA. The ability to form 2-5A structures with double stranded RNA makes this system particularly useful for inhibition of viral replication.

[0116] (2′-5′) oligoadenylate structures can be covalently linked to antisense molecules to form chimeric oligonucleotides capable of RNA cleavage (Torrence, supra). These molecules putatively bind and activate a 2-5A dependent RNase, the oligonucleotide/enzyme complex then binds to a target RNA molecule which can then be cleaved by the RNase enzyme.

[0117] Enzymatic Nucleic Acid: Several varieties of naturally-occurring enzymatic RNAs are presently known. In addition, several in vitro selection (evolution) strategies (Orgel, 1979, Proc. R. Soc. London, B 205, 435) have been used to evolve new nucleic acid catalysts capable of catalyzing cleavage and ligation of phosphodiester linkages (Joyce, 1989, Gene, 82, 83-87; Beaudry et al., 1992, Science 257, 635-641; Joyce, 1992, Scientific American 267, 90-97; Breaker et al., 1994, TIBTECH 12, 268; Bartel et al., 1993, Science 261:1411-1418; Szostak, 1993, TIBS 17, 89-93; Kumar et al., 1995, FASEB J, 9, 1183; Breaker, 1996, Curr. Op. Biotech., 7, 442; Santoro et al., 1997, Proc. Natl. Acad. Sci., 94, 4262; Tang et al., 1997, RNA 3, 914; Nakamaye & Eckstein, 1994, supra; Long & Uhlenbeck, 1994, supra; Ishizaka et al., 1995, supra; Vaish et al., 1997, Biochemistry 36, 6495; all of these are incorporated by reference herein). Each can catalyze a series of reactions including the hydrolysis of phosphodiester bonds in trans (and thus can cleave other RNA molecules) under physiological conditions.

[0118] The enzymatic nature of an enzymatic nucleic acid molecule has significant advantages, one advantage being that the concentration of enzymatic nucleic acid molecule necessary to affect a therapeutic treatment is lower. This advantage reflects the ability of the enzymatic nucleic acid molecule to act enzymatically. Thus, a single enzymatic nucleic acid molecule is able to cleave many molecules of target RNA. In addition, the enzymatic nucleic acid molecule is a highly specific inhibitor, with the specificity of inhibition depending not only on the base-pairing mechanism of binding to the target RNA, but also on the mechanism of target RNA cleavage. Single mismatches, or base-substitutions, near the site of cleavage can be chosen to completely eliminate catalytic activity of a enzymatic nucleic acid molecule.

[0119] Nucleic acid molecules having an endonuclease enzymatic activity are able to repeatedly cleave other separate RNA molecules in a nucleotide base sequence-specific manner. With the proper design, such enzymatic nucleic acid molecules can be targeted to RNA transcripts, and achieve efficient cleavage in vitro (Zaug et al., 324, Nature 429 1986; Uhlenbeck, 1987 Nature 328, 596; Kim et al., 84 Proc. Natl. Acad. Sci. USA 8788, 1987; Dreyfus, 1988, Einstein Quart. J. Bio. Med., 6, 92; Haseloff and Gerlach, 334 Nature 585, 1988; Cech, 260 JAMA 3030, 1988; and Jefferies et al., 17 Nucleic Acids Research 1371, 1989; Santoro et al., 1997 supra).

[0120] Because of their sequence specificity, trans-cleaving enzymatic nucleic acid molecules can be used as therapeutic agents for human disease (Usman & McSwiggen, 1995 Ann. Rep. Med. Chem. 30, 285-294; Christoffersen and Marr, 1995 J. Med. Chem. 38, 2023-2037). Enzymatic nucleic acid molecules can be designed to cleave specific RNA targets within the background of cellular RNA. Such a cleavage event renders the RNA non-functional and abrogates protein expression from that RNA. In this manner, synthesis of a protein associated with a disease state can be selectively inhibited (Warashina et al., 1999, Chemistry and Biology, 6, 237-250).

[0121] Enzymatic nucleic acid molecules of the invention that are allosterically regulated (“allozymes”) can be used to down-regulate PTGDS and/or PTGDR expression. These allosteric enzymatic nucleic acids or allozymes (see for example George et al., U.S. Pat. Nos. 5,834,186 and 5,741,679, Shih et al., U.S. Pat. No. 5,589,332, Nathan et al., U.S. Pat. No. 5,871,914, Nathan and Ellington, International PCT publication No. WO 00/24931, Breaker et al., International PCT Publication Nos. WO 00/26226 and 98/27104, and Sullenger et al., International PCT publication No. WO 99/29842) are designed to respond to a signaling agent, for example, mutant PTGDS and/or PTGDR protein, wild-type PTGDS and/or PTGDR protein, mutant PTGDS and/or PTGDR RNA, wild-type PTGDS and/or PTGDR RNA, other proteins and/or RNAs involved in PTGDS or PTGDR signal transduction, compounds, metals, polymers, molecules and/or drugs that are targeted to PTGDS and/or PTGDR expressing cells etc., which in turn modulates the activity of the enzymatic nucleic acid molecule. In response to interaction with a predetermined signaling agent, the allosteric enzymatic nucleic acid molecule's activity is activated or inhibited such that the expression of a particular target is selectively down-regulated. The target can comprise wild-type PTGDS, ADORA1 and/or PTGDR, mutant PTGDS, ADORA1 and/or PTGDR, and/or a predetermined component of the PTGDS, ADORA1 or PTGDR signal transduction pathway. In a specific example, allosteric enzymatic nucleic acid molecules that are activated by interaction with a RNA encoding a PTGDR protein are used as therapeutic agents in vivo. The presence of RNA encoding the PTGDS protein activates the allosteric enzymatic nucleic acid molecule that subsequently cleaves the RNA encoding a PTGDR protein resulting in the inhibition of PTGDR protein expression. In this manner, cells that express both PTGDS and PTGDR protein are selectively targeted.

[0122] In another non-limiting example, an allozyme can be activated by a PTGDS or PTGDR protein, peptide, or mutant polypeptide that causes the allozyme to inhibit the expression of PTGDS or PTGDR gene, by, for example, cleaving RNA encoded by PTGDS or PTGDR gene. In this non-limiting example, the allozyme acts as a decoy to inhibit the function of PTGDS or PTGDR and also inhibit the expression of PTGDS or PTGDR once activated by the PTGDS or PTGDR protein.

[0123] Target Sites

[0124] Targets for useful enzymatic nucleic acid molecules and antisense nucleic acids can be determined as disclosed in Draper et al., WO 93/23569; Sullivan et al, WO 93/23057; Thompson et al., WO 94/02595; Draper et al., WO 95/04818; McSwiggen et al., U.S. Pat. No. 5,525,468, and hereby incorporated by reference herein in totality. Other examples include the following PCT applications, which concern inactivation of expression of disease-related genes: WO 95/23225, WO 95/13380, WO 94/02595, incorporated by reference herein. Rather than repeat the guidance provided in those documents here, below are provided specific examples of such methods, not limiting to those in the art. Enzymatic nucleic acid molecules and antisense to such targets are designed as described in those applications and synthesized to be tested in vitro and in vivo, as also described. The sequences of human PTGDR RNAs were screened for optimal enzymatic nucleic acid and antisense target sites using a computer-folding algorithm. Antisense, hammerhead, DNAzyme, NCH, amberzyme, zinzyme, or G-Cleaver enzymatic nucleic acid molecule binding/cleavage sites were identified. These sites are shown in Tables III to VII (all sequences are 5′ to 3′ in the tables; underlined regions can be any sequence “X” or linker X, the actual sequence is not relevant here). The nucleotide base position is noted in the Tables as that site to be cleaved by the designated type of enzymatic nucleic acid molecule. While human sequences can be screened and enzymatic nucleic acid molecule and/or antisense thereafter designed, as discussed in Stinchcomb et al., WO 95/23225, mouse targeted enzymatic nucleic acid molecules can be useful to test efficacy of action of the enzymatic nucleic acid molecule and/or antisense prior to testing in humans.

[0125] Antisense, hammerhead, DNAzyme, NCH, amberzyme, zinzyme or G-Cleaver enzymatic nucleic acid molecule binding/cleavage sites were identified. The nucleic acid molecules are individually analyzed by computer folding (Jaeger et al., 1989 Proc. Natl. Acad. Sci. USA, 86, 7706) to assess whether the sequences fold into the appropriate secondary structure. Those nucleic acid molecules with unfavorable intramolecular interactions such as between the binding arms and the catalytic core are eliminated from consideration. Varying binding arm lengths can be chosen to optimize activity.

[0126] Antisense, hammerhead, DNAzyme, NCH, amberzyme, zinzyme or G-Cleaver enzymatic nucleic acid molecule binding/cleavage sites were identified and were designed to anneal to various sites in the RNA target. The binding arms are complementary to the target site sequences described above. The nucleic acid molecules were chemically synthesized. The method of synthesis used follows the procedure for normal DNA/RNA synthesis as described below and in Usman et al., 1987 J. Am. Chem. Soc., 109, 7845; Scaringe et al., 1990 Nucleic Acids Res., 18, 5433; and Wincott et al., 1995 Nucleic Acids Res. 23, 2677-2684; Caruthers et al., 1992, Methods in Enzymology 211,3-19.

[0127] Synthesis of Nucleic acid Molecules

[0128] Synthesis of nucleic acids greater than 100 nucleotides in length is difficult using automated methods, and the therapeutic cost of such molecules is prohibitive. In this invention, small nucleic acid motifs (“small refers to nucleic acid motifs less than about 100 nucleotides in length, and in one embodiment less than about 80 nucleotides in length, and in another embodiment less than about 50 nucleotides in length; e.g., antisense oligonucleotides, hammerhead or the NCH ribozymes) can be used for exogenous delivery. The simple structure of these molecules increases the ability of the nucleic acid to invade targeted regions of RNA structure. Exemplary molecules of the instant invention are chemically synthesized, and others can similarly be synthesized.

[0129] Oligonucleotides (e.g., antisense GeneBlocs) are synthesized using protocols known in the art as described in Caruthers et al., 1992, Methods in Enzymology 211, 3-19, Thompson et al., International PCT Publication No. WO 99/54459, Wincott et al., 1995, Nucleic Acids Res. 23, 2677-2684, Wincott et al., 1997, Methods Mol. Bio., 74, 59, Brennan et al., 1998, Biotechnol Bioeng., 61, 33-45, and Brennan, U.S. Pat. No. 6,001,311. All of these references are incorporated herein by reference. The synthesis of oligonucleotides makes use of common nucleic acid protecting and coupling groups, such as dimethoxytrityl at the 5′-end, and phosphoramidites at the 3′-end. In a non-limiting example, small scale syntheses are conducted on a 394 Applied Biosystems, Inc. synthesizer using a 0.2 μmol scale protocol with a 2.5 min coupling step for 2′-O-methylated nucleotides and a 45 sec coupling step for 2′-deoxy nucleotides. Table II outlines the amounts and the contact times of the reagents used in the synthesis cycle. Alternatively, syntheses at the 0.2 μmol scale can be performed on a 96-well plate synthesizer, such as the instrument produced by Protogene (Palo Alto, Calif.) with minimal modification to the cycle. A 33-fold excess (60 μL of 0.11 M=6.6 μmol) of 2′-O-methyl phosphoramidite and a 105-fold excess of S-ethyl tetrazole (60 μL of 0.25 M=15 μmol) can be used in each coupling cycle of 2′-O-methyl residues relative to polymer-bound 5′-hydroxyl. A 22-fold excess (40 μL of 0.11 M=4.4 μmol) of deoxy phosphoramidite and a 70-fold excess of S-ethyl tetrazole (40 μL of 0.25 M=10 μmol) can be used in each coupling cycle of deoxy residues relative to polymer-bound 5′-hydroxyl. Average coupling yields on the 394 Applied Biosystems, Inc. synthesizer, determined by calorimetric quantitation of the trityl fractions, are typically 97.5-99%. Other oligonucleotide synthesis reagents for the 394 Applied Biosystems, Inc. synthesizer include; detritylation solution is 3% TCA in methylene chloride (ABI); capping is performed with 16% N-methylimidazole in THF (ABI) and 10% acetic anhydride/10% 2,6-lutidine in THF (ABI); and oxidation solution is 16.9 mM 12, 49 mM pyridine, 9% water in THF (PERSEPTIVE™). Burdick & Jackson Synthesis Grade acetonitrile is used directly from the reagent bottle. S-Ethyltetrazole solution (0.25 M in acetonitrile) is made up from the solid obtained from American International Chemical, Inc. Alternately, for the introduction of phosphorothioate linkages, Beaucage reagent (3H-1,2-Benzodithiol-3-one 1,1-dioxide, 0.05 M in acetonitrile) is used.

[0130] Deprotection of the antisense oligonucleotides is performed as follows: the polymer-bound trityl-on oligoribonucleotide is transferred to a 4 mL glass screw top vial and suspended in a solution of 40% aq. methylamine (1 mL) at 65° C. for 10 min. After cooling to −20° C., the supernatant is removed from the polymer support. The support is washed three times with 1.0 mL of EtOH:MeCN:H₂O/3:1:1, vortexed and the supernatant is then added to the first supernatant. The combined supernatants, containing the oligoribonucleotide, are dried to a white powder.

[0131] The method of synthesis used for normal RNA including certain enzymatic nucleic acid molecules follows the procedure as described in Usman et al., 1987, J. Am. Chem. Soc., 109, 7845; Scaringe et al., 1990, Nucleic Acids Res., 18, 5433; and Wincott et al., 1995, Nucleic Acids Res. 23, 2677-2684 Wincott et al., 1997, Methods Mol. Bio., 74, 59, and makes use of common nucleic acid protecting and coupling groups, such as dimethoxytrityl at the 5′-end, and phosphoramidites at the 3′-end. In a non-limiting example, small scale syntheses are conducted on a 394 Applied Biosystems, Inc. synthesizer using a 0.2 μmol scale protocol with a 7.5 min coupling step for alkylsilyl protected nucleotides and a 2.5 min coupling step for 2′-O-methylated nucleotides. Table II outlines the amounts and the contact times of the reagents used in the synthesis cycle. Alternatively, syntheses at the 0.2 μmol scale can be done on a 96-well plate synthesizer, such as the instrument produced by Protogene (Palo Alto, Calif.) with minimal modification to the cycle. A 33-fold excess (60 μL of 0.11 M=6.6 μmol) of 2′-O-methyl phosphoramidite and a 75-fold excess of S-ethyl tetrazole (60 μL of 0.25 M=15 μmol) can be used in each coupling cycle of 2′-O-methyl residues relative to polymer-bound 5′-hydroxyl. A 66-fold excess (120 μL of 0.11 M=13.2 μmol) of alkylsilyl (ribo) protected phosphoramidite and a 150-fold excess of S-ethyl tetrazole (120 μL of 0.25 M=30 μmol) can be used in each coupling cycle of ribo residues relative to polymer-bound 5′-hydroxyl. Average coupling yields on the 394 Applied Biosystems, Inc. synthesizer, determined by colorimetric quantitation of the trityl fractions, are typically 97.5-99%. Other oligonucleotide synthesis reagents for the 394 Applied Biosystems, Inc. synthesizer include; detritylation solution is 3% TCA in methylene chloride (ABI); capping is performed with 16% N-methylimidazole in THF (ABI) and 10% acetic anhydride/10% 2,6-lutidine in THF (ABI); oxidation solution is 16.9 mM 12, 49 mM pyridine, 9% water in THF (PERSEPTIVE™). Burdick & Jackson Synthesis Grade acetonitrile is used directly from the reagent bottle. S-Ethyltetrazole solution (0.25 M in acetonitrile) is made up from the solid obtained from American International Chemical, Inc. Alternately, for the introduction of phosphorothioate linkages, Beaucage reagent (3H-1,2-Benzodithiol-3-one 1,1-dioxide 0.05 M in acetonitrile) is used.

[0132] Deprotection of the RNA is performed using either a two-pot or one-pot protocol. For the two-pot protocol, the polymer-bound trityl-on oligoribonucleotide is transferred to a 4 mL glass screw top vial and suspended in a solution of 40% aq. methylamine (1 mL) at 65° C. for 10 min. After cooling to −20° C., the supernatant is removed from the polymer support. The support is washed three times with 1.0 mL of EtOH:MeCN:H2O/3:1:1, vortexed and the supernatant is then added to the first supernatant. The combined supernatants, containing the oligoribonucleotide, are dried to a white powder. The base deprotected oligoribonucleotide is resuspended in anhydrous TEA/HF/NMP solution (300 μL of a solution of 1.5 mL N-methylpyrrolidinone, 750 μL TEA and 1 mL TEA.3HF to provide a 1.4 M HF concentration) and heated to 65° C. After 1.5 h, the oligomer is quenched with 1.5 M NH₄HCO₃.

[0133] Alternatively, for the one-pot protocol, the polymer-bound trityl-on oligoribonucleotide is transferred to a 4 mL glass screw top vial and suspended in a solution of 33% ethanolic methylamine/DMSO: 1/1 (0.8 mL) at 65° C. for 15 min. The vial is brought to r.t. TEA.3HF (0.1 mL) is added and the vial is heated at 65° C. for 15 min. The sample is cooled at −20° C. and then quenched with 1.5 M NH₄HCO₃.

[0134] For purification of the trityl-on oligomers, the quenched NH₄HCO₃ solution is loaded onto a C-18 containing cartridge that had been prewashed with acetonitrile followed by 50 mM TEAA. After washing, the loaded cartridge with water, the RNA is detritylated with 0.5% TFA for 13 min. The cartridge is then washed again with water, salt exchanged with 1 M NaCl and washed with water again. The oligonucleotide is then eluted with 30% acetonitrile.

[0135] Inactive hammerhead ribozymes or binding attenuated control (BAC) oligonucleotides are synthesized by substituting a U for G₅ and a U for A14 (numbering from Hertel, K. J., et al., 1992, Nucleic Acids Res., 20, 3252). Similarly, one or more nucleotide substitutions can be introduced in other enzymatic nucleic acid molecules to inactivate the molecule and such molecules can serve as a negative control.

[0136] The average stepwise coupling yields are typically >98% (Wincott et al., 1995 Nucleic Acids Res. 23, 2677-2684). Those of ordinary skill in the art will recognize that the scale of synthesis can be adapted to be larger or smaller than the example described above including but not limited to 96 well format, all that is important is the ratio of chemicals used in the reaction.

[0137] Alternatively, the nucleic acid molecules of the present invention can be synthesized separately and joined together post-synthetically, for example by ligation (Moore et al., 1992, Science 256, 9923; Draper et al., International PCT publication No. WO 93/23569; Shabarova et al., 1991, Nucleic Acids Research 19, 4247; Bellon et al., 1997, Nucleosides & Nucleotides, 16, 951; Bellon et al., 1997, Bioconjugate Chem. 8, 204).

[0138] The nucleic acid molecules of the present invention can be modified extensively to enhance stability by modification with nuclease resistant groups, for example, 2′-amino, 2′-C-allyl, 2′-flouro, 2′-O-methyl, 2′-H (for a review see Usman and Cedergren, 1992, TIBS 17, 34; Usman et al., 1994, Nucleic Acids Symp. Ser. 31, 163). Ribozymes are purified by gel electrophoresis using general methods or are purified by high pressure liquid chromatography (HPLC; See Wincott et al., Supra, the totality of which is hereby incorporated herein by reference) and are re-suspended in water.

[0139] The sequences of the nucleic acid molecules, including enzymatic nucleic acid molecules and antisense, that are chemically synthesized, are shown in Tables III-VII. The sequences of the enzymatic nucleic acid constructs that are chemically synthesized are complementary to the Substrate sequences shown in Tables III-VII. Those in the art will recognize that these sequences are representative only of many more such sequences where the enzymatic portion of the enzymatic nucleic acid (all but the binding arms) is altered to affect activity. The enzymatic nucleic acid construct sequences listed in Tables III-VII can be formed of ribonucleotides or other nucleotides or non-nucleotides. Such enzymatic nucleic acid molecules with enzymatic activity are equivalent to the enzymatic nucleic acid molecules described specifically in the Tables.

[0140] Optimizing Activity of the Nucleic Acid Molecule of the Invention.

[0141] Chemically synthesizing nucleic acid molecules with modifications (base, sugar and/or phosphate) that prevent their degradation by serum ribonucleases can increase their potency (see e.g., Eckstein et al., International Publication No. WO 92/07065; Perrault et al., 1990 Nature 344, 565; Pieken et al., 1991, Science 253, 314; Usman and Cedergren, 1992, Trends in Biochem. Sci. 17, 334; Usman et al., International Publication No. WO 93/15187; and Rossi et al., International Publication No. WO 91/03162; Sproat, U.S. Pat. No. 5,334,711; and Burgin et al., supra; all of these describe various chemical modifications that can be made to the base, phosphate and/or sugar moieties of the nucleic acid molecules herein). Modifications that enhance their efficacy in cells, and removal of bases from nucleic acid molecules to shorten oligonucleotide synthesis times and reduce chemical requirements are desired. (All these publications are hereby incorporated by reference herein).

[0142] There are several examples in the art describing sugar, base and phosphate modifications that can be introduced into nucleic acid molecules with significant enhancement in their nuclease stability and efficacy. For example, oligonucleotides are modified to enhance stability and/or enhance biological activity by modification with nuclease resistant groups, for example, 2′-amino, 2′-C-allyl, 2′-flouro, 2′-O-methyl, 2′-H, nucleotide base modifications (for a review see Usman and Cedergren, 1992, TIBS. 17, 34; Usman et al., 1994, Nucleic Acids Symp. Ser. 31, 163; Burgin et al., 1996, Biochemistry, 35, 14090). Sugar modification of nucleic acid molecules have been extensively described in the art (see Eckstein et al., International Publication PCT No. WO 92/07065; Perrault et al. Nature, 1990, 344, 565-568; Pieken et al. Science, 1991, 253, 314-317; Usman and Cedergren, Trends in Biochem. Sci., 1992, 17, 334-339; Usman et al. International Publication PCT No. WO 93/15187; Sproat, U.S. Pat. No. 5,334,711 and Beigelman et al., 1995, J. Biol. Chem., 270, 25702; Beigelman et al., International PCT publication No. WO 97/26270; Beigelman et al., U.S. Pat. No. 5,716,824; Usman et al., U.S. Pat. No. 5,627,053; Woolf et al., International PCT Publication No. WO 98/13526; Thompson et al., U.S. S No. 60/082,404 which was filed on Apr. 20, 1998; Karpeisky et al., 1998, Tetrahedron Lett., 39, 1131; Earnshaw and Gait, 1998, Biopolymers (Nucleic acid Sciences), 48, 39-55; Verma and Eckstein, 1998, Annu. Rev. Biochem., 67, 99-134; and Burlina et al., 1997, Bioorg. Med. Chem., 5, 1999-2010; all of the references are hereby incorporated in their totality by reference herein). Such publications describe general methods and strategies to determine the location of incorporation of sugar, base and/or phosphate modifications and the like into ribozymes without inhibiting catalysis, and are incorporated by reference herein. In view of such teachings, similar modifications can be used as described herein to modify the nucleic acid molecules of the instant invention.

[0143] While chemical modification of oligonucleotide internucleotide linkages with phosphorothioate, phosphorothioate, and/or 5′-methylphosphonate linkages improves stability, too many of these modifications can cause some toxicity. Therefore when designing nucleic acid molecules the amount of these internucleotide linkages should be minimized. The reduction in the concentration of these linkages should lower toxicity resulting in increased efficacy and higher specificity of these molecules.

[0144] Nucleic acid molecules having chemical modifications that maintain or enhance activity are provided. Such a nucleic acid is also generally more resistant to nucleases than an unmodified nucleic acid. Thus, in a cell and/or in vivo the activity may not be significantly lowered. Therapeutic nucleic acid molecules delivered exogenously are optimally stable within cells until translation of the target RNA has been inhibited long enough to reduce the levels of the undesirable protein. This period of time varies between hours to days depending upon the disease state. Clearly, nucleic acid molecules must be resistant to nucleases in order to function as effective intracellular therapeutic agents. Improvements in the chemical synthesis of RNA and DNA (Wincott et al., 1995 Nucleic Acids Res. 23, 2677; Caruthers et al., 1992, Methods in Enzymology 211,3-19 (incorporated by reference herein) have expanded the ability to modify nucleic acid molecules by introducing nucleotide modifications to enhance their nuclease stability as described above.

[0145] In one embodiment, nucleic acid molecules of the invention include one or more G-clamp nucleotides. A G-clamp nucleotide is a modified cytosine analog wherein modifications result in the ability to hydrogen bond both Watson-Crick and Hoogsteen faces of a complementary guanine within a duplex, see for example Lin and Matteucci, 1998, J. Am. Chem. Soc., 120, 8531-8532. A single G-clamp analog substation within an oligonucleotide can result in substantially enhanced helical thermal stability and mismatch discrimination when hybridized to complementary oligonucleotides. The inclusion of such nucleotides in nucleic acid molecules of the invention can enable both enhanced affinity and specificity to nucleic acid targets.

[0146] Therapeutic nucleic acid molecules (e.g., enzymatic nucleic acid molecules and antisense nucleic acid molecules) delivered exogenously are optimally stable within cells until translation of the target RNA has been inhibited long enough to reduce the levels of the undesirable protein. This period of time varies between hours to days depending upon the disease state. These nucleic acid molecules should be resistant to nucleases in order to function as effective intracellular therapeutic agents. Improvements in the chemical synthesis of nucleic acid molecules described in the instant invention and in the art have expanded the ability to modify nucleic acid molecules by introducing nucleotide modifications to enhance their nuclease stability as described above.

[0147] In another embodiment, the invention features conjugates and/or complexes of nucleic acid molecules targeting PTGDS, PTGDR, and/or adenosine receptors. Compositions and conjugates are used to facilitate delivery of molecules into a biological system, such as cells. The conjugates provided by the instant invention can impart therapeutic activity by transferring therapeutic compounds across cellular membranes, altering the pharmacokinetics, and/or modulating the localization of nucleic acid molecules of the invention. The present invention encompasses the design and synthesis of novel agents for the delivery of molecules, including but not limited to small molecules, lipids, phospholipids, nucleosides, nucleotides, nucleic acids, antibodies, toxins, negatively charged polymers and other polymers, for example proteins, peptides, hormones, carbohydrates, polyethylene glycols, or polyamines, across cellular membranes. In general, the transporters described are designed to be used either individually or as part of a multi-component system, with or without degradable linkers. These compounds are expected to improve delivery and/or localization of nucleic acid molecules of the invention into a number of cell types originating from different tissues, in the presence or absence of serum (see Sullenger and Cech, U.S. Pat. No. 5,854,038). Conjugates of the molecules described herein can be attached to biologically active molecules via linkers that are biodegradable, such as biodegradable nucleic acid linker molecules.

[0148] The term “biodegradable nucleic acid linker molecule” as used herein, refers to a nucleic acid molecule that is designed as a biodegradable linker to connect one molecule to another molecule, for example, a biologically active molecule. The stability of the biodegradable nucleic acid linker molecule can be modulated by using various combinations of ribonucleotides, deoxyribonucleotides, and chemically modified nucleotides, for example 2′-O-methyl, 2′-fluoro, 2′-amino, 2′-O-amino, 2′-C-allyl, 2′-O-allyl, and other 2′-modified or base modified nucleotides. The biodegradable nucleic acid linker molecule can be a dimer, trimer, tetramer or longer nucleic acid molecule, for example an oligonucleotide of about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleotides in length, or can comprise a single nucleotide with a phosphorus based linkage, for example a phosphoramidate or phosphodiester linkage. The biodegradable nucleic acid linker molecule can also comprise nucleic acid backbone, nucleic acid sugar, or nucleic acid base modifications.

[0149] The term “biodegradable” as used herein, refers to degradation in a biological system, for example enzymatic degradation or chemical degradation.

[0150] The term “biologically active molecule” as used herein, refers to compounds or molecules that are capable of eliciting or modifying a biological response in a system. Non-limiting examples of biologically active molecules contemplated by the instant invention include therapeutically active molecules such as antibodies, hormones, antivirals, peptides, proteins, chemotherapeutics, small molecules, vitamins, co-factors, nucleosides, nucleotides, oligonucleotides, enzymatic nucleic acids, antisense nucleic acids, triplex forming oligonucleotides, 2,5-A chimeras, siRNA, dsRNA, allozymes, aptamers, decoys and analogs thereof. Biologically active molecules of the invention also include molecules capable of modulating the pharmacokinetics and/or pharmacodynamics of other biologically active molecules, for example lipids and polymers such as polyamines, polyamides, polyethylene glycol and other polyethers.

[0151] The term “phospholipid” as used herein, refers to a hydrophobic molecule comprising at least one phosphorus group. For example, a phospholipid can comprise a phosphorus containing group and saturated or unsaturated alkyl group, optionally substituted with OH, COOH, oxo, amine, or substituted or unsubstituted aryl groups.

[0152] In another embodiment, nucleic acid catalysts having chemical modifications that maintain or enhance enzymatic activity are provided. Such nucleic acids are also generally more resistant to nucleases than unmodified nucleic acid. Thus, in a cell and/or in vivo the activity of the nucleic acid may not be significantly lowered. As exemplified herein such enzymatic nucleic acids are useful in a cell and/or in vivo even if activity over all is reduced 10 fold (Burgin et al., 1996, Biochemistry, 35, 14090). Such enzymatic nucleic acids herein are said to “maintain” the enzymatic activity of an all RNA ribozyme or all DNA DNAzyme.

[0153] In another aspect the nucleic acid molecules comprise a 5′ and/or a 3′-cap structure.

[0154] By “cap structure” is meant chemical modifications, which have been incorporated at either terminus of the oligonucleotide (see for example Wincott et al., WO 97/26270, incorporated by reference herein). These terminal modifications protect the nucleic acid molecule from exonuclease degradation, and can help in delivery and/or localization within a cell. The cap can be present at the 5′-terminus (5′-cap) or at the 3′-terminus (3′-cap) or can be present on both terminus. In non-limiting examples, the 5′-cap includes inverted abasic residue (moiety), 4′,5′-methylene nucleotide; 1-(beta-D-erythrofuranosyl) nucleotide, 4′-thio nucleotide, carbocyclic nucleotide; 1,5-anhydrohexitol nucleotide; L-nucleotides; alpha-nucleotides; modified base nucleotide; phosphorodithioate linkage; threo-pentofuranosyl nucleotide; acyclic 3′,4′-seco nucleotide; acyclic 3,4-dihydroxybutyl nucleotide; acyclic 3,5-dihydroxypentyl nucleotide, 3′-3′-inverted nucleotide moiety; 3′-3′-inverted abasic moiety; 3′-2′-inverted nucleotide moiety; 3′-2′-inverted abasic moiety; 1,4-butanediol phosphate; 3′-phosphoramidate; hexylphosphate; aminohexyl phosphate; 3′-phosphate; 3′-phosphorothioate; phosphorodithioate; or bridging or non-bridging methylphosphonate moiety (for more details see Wincott et al., International PCT publication No. WO 97/26270, incorporated by reference herein).

[0155] In another embodiment the 3′-cap includes, for example 4′,5′-methylene nucleotide; 1-(beta-D-erythrofuranosyl) nucleotide; 4′-thio nucleotide, carbocyclic nucleotide; 5′-amino-alkyl phosphate; 1,3-diamino-2-propyl phosphate, 3-aminopropyl phosphate; 6-aminohexyl phosphate; 1,2-aminododecyl phosphate; hydroxypropyl phosphate; 1,5-anhydrohexitol nucleotide; L-nucleotide; alpha-nucleotide; modified base nucleotide; phosphorodithioate; threo-pentofuranosyl nucleotide; acyclic 3′,4′-seco nucleotide; 3,4-dihydroxybutyl nucleotide; 3,5-dihydroxypentyl nucleotide, 5′-5′-inverted nucleotide moiety; 5′-5′-inverted abasic moiety; 5′-phosphoramidate; 5′-phosphorothioate; 1,4-butanediol phosphate; 5′-amino; bridging and/or non-bridging 5′-phosphoramidate, phosphorothioate and/or phosphorodithioate, bridging or non bridging methylphosphonate and 5′-mercapto moieties (for more details see Beaucage and Iyer, 1993, Tetrahedron 49, 1925; incorporated by reference herein).

[0156] By the term “non-nucleotide” is meant any group or compound thatcan be incorporated into a nucleic acid chain in the place of one or more nucleotide units, including either sugar and/or phosphate substitutions, and allows the remaining bases to exhibit their enzymatic activity. The group or compound is abasic in that it does not contain a commonly recognized nucleotide base, such as adenosine, guanine, cytosine, uracil or thymine.

[0157] An “alkyl” group refers to a saturated aliphatic hydrocarbon, including straight-chain, branched-chain, and cyclic alkyl groups. The alkyl group can have, for example, 1 to 12 carbons. In one embodiment of the invention, the alkyl group is a lower alkyl of from 1 to 7 carbons. In another embodiment the alkyl group is 1 to 4 carbons. The alkyl group can be substituted or unsubstituted. When substituted the substituted group(s) can be hydroxyl, cyano, alkoxy, ═O, ═S, NO₂ or N(CH₃)₂, amino, or SH. The term also includes alkenyl groups which are unsaturated hydrocarbon groups containing at least one carbon-carbon double bond, including straight-chain, branched-chain, and cyclic groups. The alkenyl group can have, for example, 1 to 12 carbons. In one embodiment of the invention the alkenyl group can be a lower alkenyl of from 1 to 7 carbons. In another embodiment the alkenyl group can be 1 to 4 carbons. The alkenyl group can be substituted or unsubstituted. When substituted the substituted group(s) can be, for example, hydroxyl, cyano, alkoxy, ═O, ═S, NO₂, halogen, N(CH₃)₂, amino, or SH. The term “alkyl” also includes alkynyl groups which have an unsaturated hydrocarbon group containing at least one carbon-carbon triple bond, including straight-chain, branched-chain, and cyclic groups. The alkynyl group can have, for example, 1 to 12 carbons. In one embodiment of the invention, the alkynyl group is a lower alkynyl of from 1 to 7 carbons. In another embodiment of the invention, the alkynyl group is 1 to 4 carbons. The alkynyl group can be substituted or unsubstituted. When substituted the substituted group(s) can be, for example, hydroxyl, cyano, alkoxy, ═O, ═S, NO₂ or N(CH₃)₂, amino or SH.

[0158] Such alkyl groups can also include aryl, alkylaryl, carbocyclic aryl, heterocyclic aryl, amide and ester groups. An “aryl” group refers to an aromatic group which has at least one ring having a conjugated p electron system and includes carbocyclic aryl, heterocyclic aryl and biaryl groups, all of which can be optionally substituted. The preferred substituent(s) of aryl groups are halogen, trihalomethyl, hydroxyl, SH, OH, cyano, alkoxy, alkyl, alkenyl, alkynyl, and amino groups. An “alkylaryl” group refers to an alkyl group (as described above) covalently joined to an aryl group (as described above). Carbocyclic aryl groups are groups wherein the ring atoms on the aromatic ring are all carbon atoms. The carbon atoms are optionally substituted. Heterocyclic aryl groups are groups having from 1 to 3 heteroatoms as ring atoms in the aromatic ring and the remainder of the ring atoms are carbon atoms. Suitable heteroatoms include oxygen, sulfur, and nitrogen, and include furanyl, thienyl, pyridyl, pyrrolyl, N-lower alkyl pyrrolo, pyrimidyl, pyrazinyl, imidazolyl and the like, all optionally substituted. An “amide” refers to an —C(O)—NH—R, where R is either alkyl, aryl, alkylaryl or hydrogen. An “ester” refers to an —C(O)—OR′, where R is either alkyl, aryl, alkylaryl or hydrogen.

[0159] By “nucleotide” is meant a heterocyclic nitrogenous base in N-glycosidic linkage with a phosphorylated sugar. Nucleotides are recognized in the art to include natural bases (standard), and modified bases well known in the art. Such bases are generally located at the 1′ position of a nucleotide sugar moiety. Nucleotides generally comprise a base, sugar and a phosphate group. The nucleotides can be unmodified or modified at the sugar, phosphate and/or base moiety, (also referred to interchangeably as nucleotide analogs, modified nucleotides, non-natural nucleotides, non-standard nucleotides and other; see for example, Usman and McSwiggen, supra; Eckstein et al., International PCT Publication No. WO 92/07065; Usman et al., International PCT Publication No. WO 93/15187; Uhlman & Peyman, supra all are hereby incorporated by reference herein). There are several examples of modified nucleic acid bases known in the art as summarized by Limbach et al., 1994, Nucleic Acids Res. 22, 2183. Some of the non-limiting examples of chemically modified and other natural nucleic acid bases that can be introduced into nucleic acids include, for example, inosine, purine, pyridin-4-one, pyridin-2-one, phenyl, pseudouracil, 2, 4, 6-trimethoxy benzene, 3-methyl uracil, dihydrouridine, naphthyl, aminophenyl, 5-alkylcytidines (e.g., 5-methylcytidine), 5-alkyluridines (e.g., ribothymidine), 5-halouridine (e.g., 5-bromouridine) or 6-azapyrimidines or 6-alkylpyrimidines (e.g. 6-methyluridine), propyne, quesosine, 2-thiouridine, 4-thiouridine, wybutosine, wybutoxosine, 4-acetylcytidine, 5-(carboxyhydroxymethyl)uridine, 5′-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluridine, beta-D-galactosylqueosine, 1-methyladenosine, 1-methylinosine, 2,2-dimethylguanosine, 3-methylcytidine, 2-methyladenosine, 2-methylguanosine, N6-methyladenosine, 7-methylguanosine, 5-methoxyaminomethyl-2-thiouridine, 5-methylaminomethyluridine, 5-methylcarbonylmethyluridine, 5-methyloxyuridine, 5-methyl-2-thiouridine, 2-methylthio-N6-isopentenyladenosine, beta-D-mannosylqueosine, uridine-5-oxyacetic acid, 2-thiocytidine, threonine derivatives and others (Burgin et al., 1996, Biochemistry, 35, 14090; Uhlman & Peyman, supra). By “modified bases” in this aspect is meant nucleotide bases other than adenine, guanine, cytosine and uracil at 1′ position or their equivalents; such bases can be used at any position, for example, within the catalytic core of an enzymatic nucleic acid molecule and/or in the substrate-binding regions of the nucleic acid molecule.

[0160] By “nucleoside” is meant a heterocyclic nitrogenous base in N-glycosidic linkage with a sugar. Nucleosides are recognized in the art to include natural bases (standard), and modified bases well known in the art. Such bases are generally located at the 1′ position of a nucleoside sugar moiety. Nucleosides generally comprise a base and sugar group. The nucleosides can be unmodified or modified at the sugar, and/or base moiety, (also referred to interchangeably as nucleoside analogs, modified nucleosides, non-natural nucleosides, non-standard nucleosides and other; see for example, Usman and McSwiggen, supra; Eckstein et al., International PCT Publication No. WO 92/07065; Usman et al., International PCT Publication No. WO 93/15187; Uhlman & Peyman, supra all are hereby incorporated by reference herein). There are several examples of modified nucleic acid bases known in the art as summarized by Limbach et al., 1994, Nucleic Acids Res. 22, 2183. Some of the non-limiting examples of chemically modified and other natural nucleic acid bases that can be introduced into nucleic acids include, inosine, purine, pyridin-4-one, pyridin-2-one, phenyl, pseudouracil, 2, 4, 6-trimethoxy benzene, 3-methyl uracil, dihydrouridine, naphthyl, aminophenyl, 5-alkylcytidines (e.g., 5-methylcytidine), 5-alkyluridines (e.g., ribothymidine), 5-halouridine (e.g., 5-bromouridine) or 6-azapyrimidines or 6-alkylpyrimidines (e.g. 6-methyluridine), propyne, quesosine, 2-thiouridine, 4-thiouridine, wybutosine, wybutoxosine, 4-acetylcytidine, 5-(carboxyhydroxymethyl)uridine, 5′-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluridine, beta-D-galactosylqueosine, 1-methyladenosine, 1-methylinosine, 2,2-dimethylguanosine, 3-methylcytidine, 2-methyladenosine, 2-methylguanosine, N6-methyladenosine, 7-methylguanosine, 5-methoxyaminomethyl-2-thiouridine, 5-methylaminomethyluridine, 5-methylcarbonylmethyluridine, 5-methyloxyuridine, 5-methyl-2-thiouridine, 2-methylthio-N6-isopentenyladenosine, beta-D-mannosylqueosine, uridine-5-oxyacetic acid, 2-thiocytidine, threonine derivatives and others (Burgin et al., 1996, Biochemistry, 35, 14090; Uhlman & Peyman, supra). By “modified bases” in this aspect is meant nucleoside bases other than adenine, guanine, cytosine and uracil at 1′ position or their equivalents; such bases can be used at any position, for example, within the catalytic core of an enzymatic nucleic acid molecule and/or in the substrate-binding regions of the nucleic acid molecule.

[0161] In one embodiment, the invention features modified enzymatic nucleic acid molecules with phosphate backbone modifications comprising one or more phosphorothioate, phosphorodithioate, methylphosphonate, morpholino, amidate carbamate, carboxymethyl, acetamidate, polyamide, sulfonate, sulfonamide, sulfamate, formacetal, thioformacetal, and/or alkylsilyl, substitutions. For a review of oligonucleotide backbone modifications see Hunziker and Leumann, 1995, Nucleic Acid Analogues: Synthesis and Properties, in Modern Synthetic Methods, VCH, 331-417, and Mesmaeker et al., 1994, Novel Backbone Replacements for Oligonucleotides, in Carbohydrate Modifications in Antisense Research, ACS, 24-39. These references are hereby incorporated by reference herein.

[0162] By “abasic” is meant sugar moieties lacking a base or having other chemical groups in place of a base at the 1′ position, for example a 3′,3′-linked or 5′,5′-linked deoxyabasic ribose derivative (for more details see Wincott et al., International PCT publication No. WO 97/26270).

[0163] By “unmodified nucleoside” is meant one of the bases adenine, cytosine, guanine, thymine, uracil joined to the 1′ carbon of β-D-ribo-furanose.

[0164] By “modified nucleoside” is meant any nucleotide base that contains a modification in the chemical structure of an unmodified nucleotide base, sugar and/or phosphate. In connection with 2′-modified nucleotides as described for the present invention, by “amino” is meant 2′-NH₂ or 2′-O-NH₂, which can be modified or unmodified. Such modified groups are described, for example, in Eckstein et al., U.S. Pat. No. 5,672,695 and Matulic-Adamic et al., WO 98/28317, respectively, which are both incorporated by reference in their entireties.

[0165] Various modifications to nucleic acid (e.g., antisense and ribozyme) structure can be made to enhance the utility of these molecules. For example, such modifications can enhance shelf-life, half-life in vitro, stability, and ease of introduction of such oligonucleotides to the target site, including e.g., enhancing penetration of cellular membranes and conferring the ability to recognize and bind to targeted cells.

[0166] Use of the nucleic acid-based molecules of the invention can lead to better treatment of the disease progression by affording the possibility of combination therapies (e.g., multiple enzymatic nucleic acid molecules targeted to different genes, enzymatic nucleic acid molecules coupled with known small molecule inhibitors, or intermittent treatment with combinations of enzymatic nucleic acid molecules (including different enzymatic nucleic acid molecule motifs) and/or other chemical or biological molecules). The treatment of patients with nucleic acid molecules can also include combinations of different types of nucleic acid molecules. Therapies can be devised which include a mixture of enzymatic nucleic acid molecules (including different enzymatic nucleic acid molecule motifs), antisense and/or 2-5A chimera molecules to one or more targets to alleviate symptoms of a disease.

[0167] Administration of Nucleic Acid Molecules

[0168] A nucleid acid molecule of the invention can be adapted for use to treat asthma and other related diseases and conditions described herein. For example, a nucleic acid molecule can comprise a delivery vehicle, including liposomes, for administration to a subject, carriers and diluents and their salts, and/or can be present in pharmaceutically acceptable formulations. Methods for the delivery of nucleic acid molecules are described in Akhtar et al., 1992, Trends Cell Bio., 2, 139; and Delivery Strategies for Antisense Oligonucleotide Therapeutics, ed. Akhtar, 1995 which are both incorporated herein by reference. Sullivan et al., PCT WO 94/02595, further describes the general methods for delivery of enzymatic RNA molecules. These protocols can be utilized for the delivery of virtually any nucleic acid molecule. Nucleic acid molecules can be administered to cells by a variety of methods known to those familiar to the art, including, but not restricted to, encapsulation in liposomes, by iontophoresis, or by incorporation into other vehicles, such as hydrogels, cyclodextrins, biodegradable nanocapsules, and bioadhesive microspheres. The nucleic acid molecules or the invention are administered via pulmonary delivery, such as by inhalation of an aerosol or spray dried formulation administered by an inhalation device or nebulizer. Alternatively, the nucleic acid/vehicle combination is locally delivered by direct injection or by use of an infusion pump. Other routes of delivery include, but are not limited to oral (tablet or pill form) and/or intrathecal delivery (Gold, 1997, Neuroscience, 76, 1153-1158). Other approaches include the use of various transport and carrier systems, for example though the use of conjugates and biodegradable polymers. For a comprehensive review on drug delivery strategies including CNS delivery, see Ho et al., 1999, Curr. Opin. Mol. Ther., 1, 336-343 and Jain, Drug Delivery Systems: Technologies and Commercial Opportunities, Decision Resources, 1998 and Groothuis et al., 1997, J NeuroVirol., 3, 387-400. More detailed descriptions of nucleic acid delivery and administration are provided in Sullivan et al., supra, Draper et al., PCT WO93/23569, Beigelman et al., PCT WO99/05094, and Klimuk et al., PCT WO99/04819 all of which have been incorporated by reference herein.

[0169] The molecules of the instant invention can be used as pharmaceutical agents. Pharmaceutical agents prevent, inhibit the occurrence, or treat (alleviate a symptom to some extent, or all of the symptoms) of a disease state in a patient.

[0170] The negatively charged polynucleotides of the invention can be administered (e.g., RNA, DNA or protein) and introduced into a patient by any standard means, with or without stabilizers, buffers, and the like, to form a pharmaceutical composition. When it is desired to use a liposome delivery mechanism, standard protocols for formation of liposomes can be followed. The compositions of the present invention can also be formulated and used as tablets, capsules or elixirs for oral administration; suppositories for rectal administration; sterile solutions; suspensions for injectable administration; and the other compositions known in the art.

[0171] The present invention also includes pharmaceutically acceptable formulations of the compounds described. These formulations include salts of the above compounds, e.g., acid addition salts, for example, salts of hydrochloric, hydrobromic, acetic acid, and benzene sulfonic acid.

[0172] A pharmacological composition or formulation refers to a composition or formulation in a form suitable for administration, e.g., local administration or systemic administration, into a cell or patient, including, for example, a human. Suitable forms, in part, depend upon the use or the route of entry, for example oral, transdermal, or by injection. Such forms should not prevent the composition or formulation from reaching a target cell (i.e., a cell to which the negatively charged polymer is desired to be delivered to). For example, pharmacological compositions injected into the blood stream should be soluble. Other factors are known in the art, and include considerations such as toxicity and forms which prevent the composition or formulation from exerting its effect.

[0173] By “local administration” is meant in vivo local absorption or accumulation of drugs in the specific tissue, organ, or compartment of the body. Administration routes that can lead to local absorption include, without limitations: inhalation, direct injection, or dermatological applications.

[0174] By “systemic administration” is meant in vivo systemic absorption or accumulation of drugs in the blood stream followed by distribution throughout the entire body. Administration routes which lead to systemic absorption include, without limitations: intravenous, subcutaneous, intraperitoneal, inhalation, oral, intrapulmonary and intramuscular. Each of these administration routes expose the desired compound, e.g., nucleic acids, to an accessible diseased tissue. The rate of entry of a drug into the circulation has been shown to be a function of molecular weight or size. The use of a liposome or other drug carrier comprising the compounds of the instant invention, for example PEG or phospholipids conjugates, can potentially localize the drug, for example, in certain tissue types, such as the tissues of the reticular endothelial system (RES). A nucleic acid formulation that can facilitate the association of drug with the surface of cells, such as, lymphocytes and macrophages is also useful. This approach can provide enhanced delivery of the drug to target cells by taking advantage of the specificity of macrophage and lymphocyte immune recognition of abnormal cells.

[0175] Both local and systemic administration approaches can be used to administer nucleic acid molecules of the invention for the treatment of asthma or related conditions. In one embodiment, the nucleic acid molecule or formulation comprising the nucleic acid molecule is administered to a patient with an inhaler or nebulizer, providing rapid local uptake of the nucleic acid molecules into relevant pulmonary tissues. In another embodiment, the nucleic acid molecule or formulation comprising the nucleic acid molecule is administered to a patient systemically, for example by intravenous or subcutaneous injection, providing sustained uptake of the nucleic acid molecules into relevant bodily tissues.

[0176] By pharmaceutically acceptable formulation is meant, a composition or formulation that allows for the effective distribution of the nucleic acid molecules of the instant invention in the physical location most suitable for their desired activity. Non-limiting examples of agents suitable for formulation with the nucleic acid molecules of the instant invention include: PEG conjugated nucleic acids, phospholipid conjugated nucleic acids, nucleic acids containing lipophilic moieties, phosphorothioates, P-glycoprotein inhibitors (such as Pluronic P85) which can enhance entry of drugs into various tissues, for exaple the CNS (Jolliet-Riant and Tillement, 1999, Fundam. Clin. Pharmacol., 13, 16-26); biodegradable polymers, such as poly (DL-lactide-coglycolide) microspheres for sustained release delivery after implantation (Emerich, DF et al, 1999, Cell Transplant, 8, 47-58) Alkermes, Inc. Cambridge, Mass.; and loaded nanoparticles, such as those made of polybutylcyanoacrylate, which can deliver drugs across the blood brain barrier and can alter neuronal uptake mechanisms (Prog Neuropsychopharmacol Biol Psychiatry, 23, 941-949, 1999). Other non-limiting examples of delivery strategies, including CNS delivery of the nucleic acid molecules of the instant invention include material described in Boado et al., 1998, J. Pharm. Sci., 87, 1308-1315; Tyler et al., 1999, FEBS Lett., 421, 280-284; Pardridge et al., 1995, PNAS USA., 92, 5592-5596; Boado, 1995, Adv. Drug Delivery Rev., 15, 73-107; Aldrian-Herrada et al., 1998, Nucleic Acids Res., 26, 4910-4916; and Tyler et al., 1999, PNAS USA., 96, 7053-7058. All these references are hereby incorporated herein by reference.

[0177] The invention also features the use of the composition comprising surface-modified liposomes containing poly (ethylene glycol) lipids (PEG-modified, or long-circulating liposomes or stealth liposomes). Nucleic acid molecules of the invention can also comprise covalently attached PEG molecules of various molecular weights. These formulations offer a method for increasing the accumulation of drugs in target tissues. This class of drug carriers resists opsonization and elimination by the mononuclear phagocytic system (MPS or RES), thereby enabling longer blood circulation times and enhanced tissue exposure for the encapsulated drug (Lasic et al. Chem. Rev. 1995, 95, 2601-2627; Ishiwata et al., Chem. Pharm. Bull. 1995, 43, 1005-1011). Such liposomes have been shown to accumulate selectively in tumors, presumably by extravasation and capture in the neovascularized target tissues (Lasic et al., Science 1995, 267, 1275-1276; Oku et al., 1995, Biochim. Biophys. Acta, 1238, 86-90). The long-circulating liposomes enhance the pharmacokinetics and pharmacodynamics of DNA and RNA, particularly compared to conventional cationic liposomes which are known to accumulate in tissues of the MPS (Liu et al., J. Biol. Chem. 1995, 42, 24864-24870; Choi et al., International PCT Publication No. WO 96/10391; Ansell et al., International PCT Publication No. WO 96/10390; Holland et al., International PCT Publication No. WO 96/10392; all of which are incorporated by reference herein). Long-circulating liposomes are also likely to protect drugs from nuclease degradation to a greater extent compared to cationic liposomes, based on their ability to avoid accumulation in metabolically aggressive MPS tissues such as the liver and spleen. All of these references are incorporated by reference herein.

[0178] The present invention also includes compositions prepared for storage or administration that include a pharmaceutically effective amount of the desired compounds in a pharmaceutically acceptable carrier or diluent. Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical art, and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (A. R. Gennaro edit. 1985) hereby incorporated by reference herein. For example, preservatives, stabilizers, dyes and flavoring agents can be provided. These include sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid. In addition, antioxidants and suspending agents can be used.

[0179] A pharmaceutically effective dose is that dose required to prevent, inhibit the occurrence, or treat (alleviate a symptom to some extent, or all of the symptoms) of a disease state. The pharmaceutically effective dose depends on the type of disease, the composition used, the route of administration, the type of mammal being treated, the physical characteristics of the specific mammal under consideration, concurrent medication, and other factors which those skilled in the medical arts will recognize. Generally, an amount between 0.1 mg/kg and 100 mg/kg body weight/day of active ingredients is administered dependent upon potency of the negatively charged polymer.

[0180] The nucleic acid molecules of the invention and formulations thereof can be administered orally, topically, parenterally, by inhalation or spray or rectally in dosage unit formulations containing conventional non-toxic pharmaceutically acceptable carriers, adjuvants and vehicles. The term parenteral as used herein includes percutaneous, subcutaneous, intravascular (e.g., intravenous), intramuscular, or intrathecal injection or infusion techniques and the like. In addition, there is provided a pharmaceutical formulation comprising a nucleic acid molecule of the invention and a pharmaceutically acceptable carrier. One or more nucleic acid molecules of the invention can be present in association with one or more non-toxic pharmaceutically acceptable carriers and/or diluents and/or adjuvants, and if desired other active ingredients. The pharmaceutical compositions containing nucleic acid molecules of the invention can be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion, hard or soft capsules, or syrups or elixirs.

[0181] Compositions intended for oral use can be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions can contain one or more such sweetening agents, flavoring agents, coloring agents or preservative agents in order to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients that are suitable for the manufacture of tablets. These excipients can be for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example starch, gelatin or acacia, and lubricating agents, for example magnesium stearate, stearic acid or talc. The tablets can be uncoated or they can be coated by known techniques. In some cases such coatings can be prepared by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monosterate or glyceryl distearate can be employed.

[0182] Formulations for oral use can also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example peanut oil, liquid paraffin or olive oil.

[0183] Aqueous suspensions contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents, for example sodium carboxymethylcellulose, methylcellulose, hydropropyl-methylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents can be a naturally-occurring phosphatide, for example, lecithin, or condensation products of an alkylene oxide with fatty acids, for example polyoxyethylene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions can also contain one or more preservatives, for example ethyl, or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharin.

[0184] Oily suspensions can be formulated by suspending the active ingredients in a vegetable oil, for example arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions can contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents and flavoring agents can be added to provide palatable oral preparations. These compositions can be preserved by the addition of an anti-oxidant such as ascorbic acid.

[0185] Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents or suspending agents are exemplified by those already mentioned above. Additional excipients, for example sweetening, flavoring and coloring agents, can also be present.

[0186] Pharmaceutical compositions of the invention can also be in the form of oil-in-water emulsions. The oily phase can be a vegetable oil or a mineral oil or mixtures of these. Suitable emulsifying agents can be naturally-occurring gums, for example gum acacia or gum tragacanth, naturally-occurring phosphatides, for example soy bean, lecithin, and esters or partial esters derived from fatty acids and hexitol, anhydrides, for example sorbitan monooleate, and condensation products of the said partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate. The emulsions can also contain sweetening and flavoring agents.

[0187] Syrups and elixirs can be formulated with sweetening agents, for example glycerol, propylene glycol, sorbitol, glucose or sucrose. Such formulations can also contain a demulcent, a preservative and flavoring and coloring agents. The pharmaceutical compositions can be in the form of a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents that have been mentioned above. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parentally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono-or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

[0188] The nucleic acid molecules of the invention can also be administered in the form of suppositories, e.g., for rectal administration of the drug. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials include cocoa butter and polyethylene glycols.

[0189] Nucleic acid molecules of the invention can be administered parenterally in a sterile medium. The drug, depending on the vehicle and concentration used, can either be suspended or dissolved in the vehicle. Advantageously, adjuvants such as local anesthetics, preservatives and buffering agents can be dissolved in the vehicle.

[0190] Dosage levels of the order of from about 0.1 mg to about 140 mg per kilogram of body weight per day are useful in the treatment of the above-indicated conditions (about 0.5 mg to about 7 g per patient per day). The amount of active ingredient that can be combined with the carrier materials to produce a single dosage form varies depending upon the host treated and the particular mode of administration. Dosage unit forms generally contain between from about 1 mg to about 500 mg of an active ingredient.

[0191] It is understood that the specific dose level for any particular patient depends upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, and rate of excretion, drug combination and the severity of the particular disease undergoing therapy.

[0192] For administration to non-human animals, the composition can also be added to the animal feed or drinking water. It can be convenient to formulate the animal feed and drinking water compositions so that the animal takes in a therapeutically appropriate quantity of the composition along with its diet. It can also be convenient to present the composition as a premix for addition to the feed or drinking water.

[0193] The nucleic acid molecules of the present invention can also be administered to a patient in combination with other therapeutic compounds to increase the overall therapeutic effect. The use of multiple compounds to treat an indication can increase the beneficial effects while reducing the presence of side effects.

[0194] Alternatively, certain of the nucleic acid molecules of the instant invention can be expressed within cells from eukaryotic promoters (e.g., Izant and Weintraub, 1985, Science, 229, 345; McGarry and Lindquist, 1986, Proc. Natl. Acad. Sci., USA 83, 399; Scanlon et al., 1991, Proc. Natl. Acad. Sci. USA, 88, 10591-5; Kashani-Sabet et al., 1992, Antisense Res. Dev., 2, 3-15; Dropulic et al., 1992, J. Virol., 66, 1432-41; Weerasinghe et al., 1991, J. Virol., 65, 5531-4; Ojwang et al., 1992, Proc. Natl. Acad. Sci. USA, 89, 10802-6; Chen et al., 1992, Nucleic Acids Res., 20, 4581-9; Sarver et al., 1990 Science, 247, 1222-1225; Thompson et al., 1995, Nucleic Acids Res., 23, 2259; Good et al., 1997, Gene Therapy, 4, 45; all of these references are hereby incorporated in their totalities by reference herein). Those skilled in the art realize that any nucleic acid can be expressed in eukaryotic cells from the appropriate DNA/RNA vector. The activity of such nucleic acids can be augmented by their release from the primary transcript by a enzymatic nucleic acid (Draper et al, PCT WO 93/23569, and Sullivan et al., PCT WO 94/02595; Ohkawa et al., 1992, Nucleic Acids Symp. Ser., 27, 15-6; Taira et al., 1991, Nucleic Acids Res., 19, 5125-30; Ventura et al., 1993, Nucleic Acids Res., 21, 3249-55; Chowrira et al., 1994, J. Biol. Chem., 269, 25856; all of these references are hereby incorporated in their totalities by reference herein). Gene therapy approaches specific to the CNS are described by Blesch et al., 2000, Drug News Perspect., 13, 269-280; Peterson et al., 2000, Cent. Nerv. Syst. Dis., 485-508; Peel and Klein, 2000, J. Neurosci. Methods, 98, 95-104; Hagihara et al., 2000, Gene Ther., 7, 759-763; and Herrlinger et al., 2000, Methods Mol. Med., 35, 287-312. AAV-mediated delivery of nucleic acid to cells of the nervous system is further described by Kaplitt et al., U.S. Pat. No. 6,180,613.

[0195] In another aspect of the invention, RNA molecules of the present invention can be expressed from transcription units (see for example Couture et al., 1996, TIG., 12, 510) inserted into DNA or RNA vectors. The recombinant vectors can be DNA plasmids or viral vectors. Ribozyme expressing viral vectors can be constructed based on, but not limited to, adeno-associated virus, retrovirus, adenovirus, or alphavirus. The recombinant vectors capable of expressing the nucleic acid molecules can be delivered as described above, and persist in target cells. Alternatively, viral vectors can be used that provide for transient expression of nucleic acid molecules. Such vectors can be repeatedly administered as necessary. Once expressed, the nucleic acid molecule binds to the target mRNA. Delivery of nucleic acid molecule expressing vectors can be systemic, such as by intravenous or intra-muscular administration, by administration to target cells ex-planted from the patient followed by reintroduction into the patient, or by any other means that would allow for introduction into the desired target cell (for a review see Couture et al., 1996, TIG., 12, 510).

[0196] In one aspect the invention features an expression vector comprising a nucleic acid sequence encoding at least one of the nucleic acid molecules of the instant invention is disclosed. The nucleic acid sequence encoding the nucleic acid molecule of the instant invention is operable linked in a manner that allows expression of that nucleic acid molecule.

[0197] In another aspect the invention features an expression vector comprising: a) a transcription initiation region (e.g., eukaryotic pol I, II or III initiation region); b) a transcription termination region (e.g., eukaryotic pol I, II or III termination region); c) a nucleic acid sequence encoding at least one of the nucleic acid catalyst of the instant invention; and wherein said sequence is operably linked to said initiation region and said termination region, in a manner that allows expression and/or delivery of said nucleic acid molecule. The vector can optionally include an open reading frame (ORF) for a protein operably linked on the 5′ side or the 3′-side of the sequence encoding the nucleic acid catalyst of the invention; and/or an intron (intervening sequences).

[0198] Transcription of the nucleic acid molecule sequences are driven from a promoter for eukaryotic RNA polymerase I (pol I), RNA polymerase II (pol II), or RNA polymerase III (pol III). Transcripts from pol II or pol III promoters are expressed at high levels in all cells; the levels of a given pol II promoter in a given cell type depends on the nature of the gene regulatory sequences (enhancers, silencers, etc.) present nearby. Prokaryotic RNA polymerase promoters are also used, providing that the prokaryotic RNA polymerase enzyme is expressed in the appropriate cells (Elroy-Stein and Moss, 1990, Proc. Natl. Acad. Sci. USA, 87, 6743-7; Gao and Huang 1993, Nucleic Acids Res., 21, 2867-72; Lieber et al., 1993, Methods Enzymol., 217, 47-66; Zhou et al., 1990, Mol. Cell. Biol., 10, 4529-37). All of these references are incorporated by reference herein. Several investigators have demonstrated that nucleic acid molecules, such as ribozymes expressed from such promoters can function in mammalian cells (e.g. Kashani-Sabet et al., 1992, Antisense Res. Dev., 2, 3-15; Ojwang et al., 1992, Proc. Natl. Acad. Sci. USA, 89, 10802-6; Chen et al., 1992, Nucleic Acids Res., 20, 4581-9; Yu et al., 1993, Proc. Natl. Acad. Sci. USA, 90, 6340-4; L'Huillier et al., 1992, EMBO J, 11, 4411-8; Lisziewicz et al., 1993, Proc. Natl. Acad. Sci. U.S. A, 90, 8000-4; Thompson et al., 1995, Nucleic Acids Res., 23, 2259; Sullenger & Cech, 1993, Science, 262, 1566). More specifically, transcription units such as the ones derived from genes encoding U6 small nuclear (snRNA), transfer RNA (tRNA) and adenovirus VA RNA are useful in generating high concentrations of desired RNA molecules such as ribozymes in cells (Thompson et al., supra; Couture and Stinchcomb, 1996, supra; Noonberg et al., 1994, Nucleic Acid Res., 22, 2830; Noonberg et al., U.S. Pat. No. 5,624,803; Good et al., 1997, Gene Ther., 4, 45; Beigelman et al., International PCT Publication No. WO 96/18736; all of these publications are incorporated by reference herein). The above ribozyme transcription units can be incorporated into a variety of vectors for introduction into mammalian cells, including but not restricted to, plasmid DNA vectors, viral DNA vectors (such as adenovirus or adeno-associated virus vectors), or viral RNA vectors (such as retroviral or alphavirus vectors) (for a review see Couture and Stinchcomb, 1996, supra).

[0199] In another aspect the invention features an expression vector comprising a nucleic acid sequence encoding at least one of the nucleic acid molecules of the invention, in a manner that allows expression of that nucleic acid molecule. The expression vector comprises in one embodiment; a) a transcription initiation region; b) a transcription termination region; c) a nucleic acid sequence encoding at least one said nucleic acid molecule; and wherein said sequence is operably linked to said initiation region and said termination region, in a manner that allows expression and/or delivery of said nucleic acid molecule.

[0200] In another embodiment the expression vector comprises: a) a transcription initiation region; b) a transcription termination region; c) an open reading frame; d) a nucleic acid sequence encoding at least one said nucleic acid molecule, wherein said sequence is operably linked to the 3′-end of said open reading frame; and wherein said sequence is operably linked to said initiation region, said open reading frame and said termination region, in a manner that allows expression and/or delivery of said nucleic acid molecule. In yet another embodiment the expression vector comprises: a) a transcription initiation region; b) a transcription termination region; c) an intron; d) a nucleic acid sequence encoding at least one said nucleic acid molecule; and wherein said sequence is operably linked to said initiation region, said intron and said termination region, in a manner which allows expression and/or delivery of said nucleic acid molecule.

[0201] In another embodiment, the expression vector comprises: a) a transcription initiation region; b) a transcription termination region; c) an intron; d) an open reading frame; e) a nucleic acid sequence encoding at least one said nucleic acid molecule, wherein said sequence is operably linked to the 3′-end of said open reading frame; and wherein said sequence is operably linked to said initiation region, said intron, said open reading frame and said termination region, in a manner which allows expression and/or delivery of said nucleic acid molecule.

EXAMPLES

[0202] The following are non-limiting examples showing the selection, isolation, synthesis and activity of nucleic acids of the instant invention.

[0203] The following examples demonstrate the selection and design of Antisense, hammerhead, DNAzyme, NCH, Amberzyme, Zinzyme, or G-Cleaver ribozyme molecules and binding/cleavage sites within PTGDS and/or PTGDR RNA.

Example 1 Identification of Potential Target Sites in Human PTGDS, ADORA1 and PTGDR RNA

[0204] The sequence of human PTGDS, ADORA1 and PTGDR genes are screened for accessible sites using a computer-folding algorithm. Regions of the RNA that do not form secondary folding structures and contained potential enzymatic nucleic acid molecule and/or antisense binding/cleavage sites are identified. The sequences of PTGDR binding/cleavage sites are shown in Tables III-VII.

Example 2 Selection of Enzymatic Nucleic Acid Cleavage Sites in Human PTGDS, ADORA1 and PTGDR RNA

[0205] Enzymatic nucleic acid molecule target sites are chosen by analyzing sequences of Human PTGDS (Genbank accession No: NM 000954), ADORA1 (Genbank accession No: NM_(—)000674) and PTGDR gene (Genbank accession Nos: U31332 and U31099) and prioritizing the sites on the basis of folding. Enzymatic nucleic acid molecules are designed that can bind each target and are individually analyzed by computer folding (Christoffersen et al., 1994 J. Mol. Struc. Theochem, 311, 273; Jaeger et al., 1989, Proc. Natl. Acad. Sci. USA, 86, 7706) to assess whether the enzymatic nucleic acid molecule sequences fold into the appropriate secondary structure. Those enzymatic nucleic acid molecules with unfavorable intramolecular interactions between the binding arms and the catalytic core are eliminated from consideration. As noted below, varying binding arm lengths can be chosen to optimize activity. Generally, at least 4 bases on each arm are able to bind to, or otherwise interact with, the target RNA.

Example 3 Chemical Synthesis and Purification of Ribozymes and Antisense for Efficient Cleavage and/or blocking of PTGDS, ADORA1 and PTGDR RNA

[0206] Enzymatic nucleic acid molecules and antisense constructs are designed to anneal to various sites in the RNA message. The binding arms of the enzymatic nucleic acid molecules are complementary to the target site sequences described above, while the antisense constructs are fully complementary to the target site sequences described above. The enzymatic nucleic acid molecules and antisense constructs were chemically synthesized. The method of synthesis used followed the procedure for normal RNA synthesis as described above and in Usman et al., (1987 J. Am. Chem. Soc., 109, 7845), Scaringe et al., (1990 Nucleic Acids Res., 18, 5433) and Wincott et al., supra, and made use of common nucleic acid protecting and coupling groups, such as dimethoxytrityl at the 5′-end, and phosphoramidites at the 3′-end. The average stepwise coupling yields were typically >98%.

[0207] Enzymatic nucleic acid molecules and antisense constructs are also synthesized from DNA templates using bacteriophage T7 RNA polymerase (Milligan and Uhlenbeck, 1989, Methods Enzymol. 180, 51). Enzymatic nucleic acid molecules and antisense constructs are purified by gel electrophoresis using general methods or are purified by high pressure liquid chromatography (HPLC; See Wincott et al., supra; the totality of which is hereby incorporated herein by reference) and are resuspended in water. The sequences of the chemically synthesized enzymatic nucleic acid molecules used in this study are shown below in Table III-VII. The sequences of the chemically synthesized antisense constructs used in this study are complementary sequences to the Substrate sequences shown below as in Table III-VII.

Example 4 Enzymatic Nucleic Acid Molecule Cleavage of PTGDS, ADORA1 and PTGDR RNA Target in vitro

[0208] Enzymatic nucleic acid molecules targeted to the human PTGDS, ADORA1 and PTGDR RNA are designed and synthesized as described above. These enzymatic nucleic acid molecules can be tested for cleavage activity in vitro, for example, using the following procedure. The target sequences and the nucleotide location within the PTGDR RNA are given in Tables III-VII.

[0209] Cleavage Reactions: Full-length or partially full-length, internally-labeled target RNA for enzymatic nucleic acid molecule cleavage assay is prepared by in vitro transcription in the presence of [a-³²P] CTP, passed over a G 50 Sephadex column by spin chromatography and used as substrate RNA without further purification. Alternately, substrates are 5′-³²P-end labeled using T4 polynucleotide kinase enzyme. Assays are performed by pre-warming a 2×concentration of purified enzymatic nucleic acid molecule in enzymatic nucleic acid molecule cleavage buffer (50 mM Tris-HCl, pH 7.5 at 37° C., 10 mM MgCl₂) and the cleavage reaction was initiated by adding the 2×enzymatic nucleic acid molecule mix to an equal volume of substrate RNA (maximum of 1-5 nM) that was also pre-warmed in cleavage buffer. As an initial screen, assays are carried out for 1 hour at 37° C. using a final concentration of either 40 nM or 1 mM enzymatic nucleic acid molecule, i.e., enzymatic nucleic acid molecule excess. The reaction is quenched by the addition of an equal volume of 95% formamide, 20 mM EDTA, 0.05% bromophenol blue and 0.05% xylene cyanol after which the sample is heated to 95° C. for 2 minutes, quick chilled and loaded onto a denaturing polyacrylamide gel. Substrate RNA and the specific RNA cleavage products generated by enzymatic nucleic acid molecule cleavage are visualized on an autoradiograph of the gel. The percentage of cleavage is determined by Phosphor Imager® quantitation of bands representing the intact substrate and the cleavage products.

Example 5 In vivo Models used to Evaluate the Down-Regulation of PTGDS, ADORA1 and PTGDR Gene Expression

[0210] Animal Models

[0211] Evaluating the efficacy of anti-PTGDS, ADORA-1 and/or PTGDR agents in animal models is an important prerequisite to human clinical trials. Matsuoka et al., 2000, Science, 287, 2012-2016, describe a useful asthma animal model having generating mice deficient in the PTGDR receptor. Sensitization and aerosol challenge of homozygous (PTGDR−/−) mice with ovalbumin was shown to induce increases in the serum concentration of immunoglobin E (IgE), an allergic mediator that activates mast cells, similar to wild-type mice subjected to the same conditions. The concentration of TH2 cytokines and the degree of lymphocyte lung infiltration in the OVA challenged PTGDR −/− mice was shown to be greatly reduced compared to wild type mice. In addition, the PTGDR −/− mice showed only marginal eosinophil infiltration and failed to develop airway hyperreactivity. Similarly, this model can be used to evaluate mice that are treated with nucleic acid molecules of the invention and can furthermore be used as a positive control in determining the response of mice treated with nucleic acid molecules of the invention by using such factors as airway obstruction, lung capacity, and bronchiolar alveolar lavage (BAL) fluid in the evaluation.

[0212] Cell Culture

[0213] Two human cell lines, NPE cells and NCB-20 cells are known to express PTGDR. Cloned human PTGDR has been expressed in CHO and COS7 cells and used in various studies. These PTGDR expressing lung cell lines can be used in cell culture assays to evaluate nucleic acid molecules of the invention. A primary endpoint in these experiments would be the RT-PCR analysis of PTGDR mRNA expression in PTGDR expressing cells. In addition, ligand binding assays can be developed where binding of PTGDS can be evaluated in response to treatment with nucleic acid molecules of the invention.

[0214] Indications

[0215] The present body of knowledge in PTGDS, ADORA1 and PTGDR research indicates the need for methods to assay PTGDS, ADORA1 and PTGDR activity and for compounds that can regulate PTGDS, ADORA1 and PTGDR expression for research, diagnostic, and therapeutic use. As described herein, the nucleic acid molecules of the present invention can be used in assays to diagnose disease state related of PTGDS, ADORA1 and/or PTGDR levels. In addition, the nucleic acid molecules can be used to treat disease state related to PTGDS, ADORA1 and/or PTGDR levels.

[0216] Particular degenerative and disease states that can be associated with PTGDS, ADORA1 and PTGDR levels include, but are not limited to allergic diseases and conditions, including but not limited to asthma, allergic rhinitis, atopic dermatitis, and any other diseases or conditions that are related to or will respond to the levels of PTGDS, ADORA1 and/or PTGDR in a cell or tissue, alone or in combination with other therapies.

[0217] The use of anti-inflammatories, bronchodilators, adenosine inhibitors and adenosine A1 receptor inhibitors are examples of other treatments or therapies can be combined with the nucleic acid molecules of the invention. Those skilled in the art will recognize that other drug compounds and therapies can be similarly be readily combined with the nucleic acid molecules of the instant invention (e.g. enzymatic nucleic acid molecules and antisense molecules) are hence within the scope of the instant invention.

[0218] Diagnostic Uses

[0219] The nucleic acid molecules of this invention (e.g., enzymatic nucleic acid molecules) can be used as diagnostic tools to examine genetic drift and mutations within diseased cells or to detect the presence of PTGDS, ADORA1 and/or PTGDR RNA in a cell. The close relationship between enzymatic nucleic acid molecule activity and the structure of the target RNA allows the detection of mutations in any region of the molecule that alters the base-pairing and three-dimensional structure of the target RNA. By using multiple enzymatic nucleic acid molecules described in this invention, one can map nucleotide changes which are important to RNA structure and function in vitro, as well as in cells and tissues. Cleavage of target RNAs with enzymatic nucleic acid molecules can be used to inhibit gene expression and define the role (essentially) of specified gene products in the progression of disease. In this manner, other genetic targets can be defined as important mediators of the disease. These experiments can lead to better treatment of the disease progression by affording the possibility of combinational therapies (e.g., multiple enzymatic nucleic acid molecules targeted to different genes, enzymatic nucleic acid molecules coupled with known small molecule inhibitors, or intermittent treatment with combinations of enzymatic nucleic acid molecules and/or other chemical or biological molecules). Other in vitro uses of enzymatic nucleic acid molecules of this invention are well known in the art, and include detection of the presence of mRNAs associated with PTGDS, ADORA1 or PTGDR-related condition. Such RNA is detected by determining the presence of a cleavage product after treatment with an enzymatic nucleic acid molecule using standard methodology.

[0220] In a specific example, enzymatic nucleic acid molecules which cleave only wild-type or mutant forms of the target RNA are used for the assay. The first enzymatic nucleic acid molecule is used to identify wild-type RNA present in the sample and the second enzymatic nucleic acid molecule is used to identify mutant RNA in the sample. As reaction controls, synthetic substrates of both wild-type and mutant RNA are cleaved by both enzymatic nucleic acid molecules to demonstrate the relative enzymatic nucleic acid molecule efficiencies in the reactions and the absence of cleavage of the “non-targeted” RNA species. The cleavage products from the synthetic substrates also serve to generate size markers for the analysis of wild-type and mutant RNAs in the sample population. Thus each analysis requires two enzymatic nucleic acid molecules, two substrates and one unknown sample which is combined into six reactions. The presence of cleavage products is determined using an RNAse protection assay so that full-length and cleavage fragments of each RNA can be analyzed in one lane of a polyacrylamide gel. It is not absolutely required to quantify the results to gain insight into the expression of mutant RNAs and putative risk of the desired phenotypic changes in target cells. The expression of mRNA whose protein product is implicated in the development of the phenotype (i.e., PTGDS/PTGDR) is adequate to establish risk. If probes of comparable specific activity are used for both transcripts, then a qualitative comparison of RNA levels will be adequate and will decrease the cost of the initial diagnosis. Higher mutant form to wild-type ratios are correlated with higher risk whether RNA levels are compared qualitatively or quantitatively. The use of enzymatic nucleic acid molecules in diagnostic applications contemplated by the instant invention is described, for example, in George et al., U.S. Pat. Nos. 5,834,186 and 5,741,679, Shih et al., U.S. Pat. No. 5,589,332, Nathan et al., U.S. Pat. No. 5,871,914, Nathan and Ellington, International PCT publication No. WO 00/24931, Breaker et al., International PCT Publication Nos. WO 00/26226 and 98/27104, and Sullenger et al., International PCT publication No. WO 99/29842.

[0221] Additional Uses

[0222] Potential uses of sequence-specific enzymatic nucleic acid molecules of the instant invention can have many of the same applications for the study of RNA that DNA restriction endonucleases have for the study of DNA (Nathans et al., 1975 Ann. Rev. Biochem. 44:273). For example, the pattern of restriction fragments can be used to establish sequence relationships between two related RNAs, and large RNAs can be specifically cleaved to fragments of a size more useful for study. The ability to engineer sequence specificity of the enzymatic nucleic acid molecule is ideal for cleavage of RNAs of unknown sequence. Applicant has described the use of nucleic acid molecules to down-regulate gene expression of target genes in bacterial, microbial, fungal, viral, and eukaryotic systems including plant, or mammalian cells.

[0223] All patents and publications mentioned in the specification are indicative of the levels of skill of those skilled in the art to which the invention pertains. All references cited in this disclosure are incorporated by reference to the same extent as if each reference had been incorporated by reference in its entirety individually.

[0224] One skilled in the art would readily appreciate that the present invention is well adapted to carry out the objects and obtain the ends and advantages mentioned, as well as those inherent therein. The methods and compositions described herein as presently representative of preferred embodiments are exemplary and are not intended as limitations on the scope of the invention. Changes therein and other uses will occur to those skilled in the art, which are encompassed within the spirit of the invention, are defined by the scope of the claims.

[0225] It will be readily apparent to one skilled in the art that varying substitutions and modifications may be made to the invention disclosed herein without departing from the scope and spirit of the invention. Thus, such additional embodiments are within the scope of the present invention and the following claims.

[0226] The invention illustratively described herein suitably can be practiced in the absence of any element or elements, limitation or limitations that is not specifically disclosed herein. Thus, for example, in each instance herein any of the terms “comprising”, “consisting essentially of” and “consisting of” can be replaced with either of the other two terms. The terms and expressions that have been employed are used as terms of description and not of limitation, and there is no intention that in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the invention claimed. Thus, it should be understood that although the present invention has been specifically disclosed by preferred embodiments, optional features, modification and variation of the concepts herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this invention as defined by the description and the appended claims.

[0227] In addition, where features or aspects of the invention are described in terms of Markush groups or other grouping of alternatives, those skilled in the art will recognize that the invention is also thereby described in terms of any individual member or subgroup of members of the Markush group or other group.

[0228] Other embodiments are within the following claims. TABLE I Characteristics of naturally occurring ribozymes Group I Introns Size: ˜150 to >1000 nucleotides. Requires a U in the target sequence immediately 5′ of the cleavage site. Binds 4-6 nucleotides at the 5′-side of the cleavage site. Reaction mechanism: attack by the 3′-OH of guanosine to generate cleavage products with 3′-OH and 5′-guanosine. Additional protein cofactors required in some cases to help folding and maintenance of the active structure. Over 300 known members of this class. Found as an intervening sequence in Tetrahymena thermophila rRNA, fungal mitochondria, chloroplasts, phage T4, blue- green algae, and others. Major structural features largely established though phylogenetic comparisons, mutagenesis, and biochemical studies [^(i),^(ii)]. Complete kinetic framework established for one ribozyme [^(iii),^(iv),^(v),^(vi)]. Studies of ribozyme folding and substrate docking underway [^(vii),^(viii),^(ix)]. Chemical modification investigation of important residues well established [^(x),^(xi)]. The small (4-6 nt) binding site may make this ribozyme too non-specific for targeted RNA cleavage, however, the Tetrahymena group I intron has been used to repair a “defective” β-galactosidase message by the ligation of new β- galactosidase sequences onto the defective message [^(xii)]. RNAse P RNA (M1 RNA) Size: ˜290 to 400 nucleotides. RNA portion of a ubiquitous ribonucleoprotein enzyme. Cleaves tRNA precursors to form mature tRNA [^(xiii)]. Reaction mechanism: possible attack by M²⁺-OH to generate cleavage products with 3′-OH and 5′-phosphate. RNAse P is found throughout the prokaryotes and eukaryotes. The RNA subunit has been sequenced from bacteria, yeast, rodents, and primates. Recruitment of endogenous RNAse P for therapeutic applications is possible through hybridization of an External Guide Sequence (EGS) to the target RNA [^(xiv),^(xv)] Important phosphate and 2′OH contacts recently identified [^(xvi),^(xvii)] Group II Introns Size: >1000 nucleotides. Trans cleavage of target RNAs recently demonstrated [^(xviii),^(xix)]. Sequence requirements not fully determined. Reaction mechanism: 2′-OH of an internal adenosine generates cleavage products with 3′-OH and a “lariat” RNA containing a 3′-5′ and a 2′-5′ branch point. Only natural ribozyme with demonstrated participation in DNA cleavage [^(xx),^(xxi)] in addition to RNA cleavage and ligation. Major structural features largely established through phylogenetic comparisons [^(xxii)]. Important 2′OH contacts beginning to be identified [^(xxiii)] Kinetic framework under development [^(xxiv)] Neurospora VS RNA Size: ˜144 nucleotides. Trans cleavage of hairpin target RNAs recently demonstrated [^(xxv)]. Sequence requirements not fully determined. Reaction mechanism: attack by 2′-OH 5′to the scissile bond to generate cleavage products with 2′,3′-cyclic phosphate and 5′-OH ends. Binding sites and structural requirements not fully determined. Only 1 known member of this class. Found in Neurospora VS RNA. Hammerhead Ribozyme (see text for references) Size: ˜13 to 40 nucleotides. Requires the target sequence UH immediately 5′ of the cleavage site. Binds a variable number nucleotides on both sides of the cleavage site. Reaction mechanism: attack by 2′-OH 5′ to the scissile bond to generate cleavage products with 2′,3′-cyclic phosphate and 5′-OH ends. 14 known members of this class. Found in a number of plant pathogens (virusoids) that use RNA as the infectious agent. Essential structural features largely defined, including 2 crystal structures [^(xxvi),^(xxvii)] Minimal ligation activity demonstrated (for engineering through in vitro selection) [^(xxviii)] Complete kinetic framework established for two or more ribozymes [^(xxix)]. Chemical modification investigation of important residues well established [^(xxx)]. Hairpin Ribozyme Size: ˜50 nucleotides. Requires the target sequence GUC immediately 3′of the cleavage site. Binds 4-6 nucleotides at the 5′-side of the cleavage site and a variable number to the 3′-side of the cleavage site. Reaction mechanism: attack by 2′-OH 5′ to the scissile bond to generate cleavage products with 2′,3′-cyclic phosphate and 5′-OH ends. 3 known members of this class. Found in three plant pathogen (satellite RNAs of the tobacco ringspot virus, arabis mosaic virus and chicory yellow mottle virus) which uses RNA as the infectious agent. Essential structural features largely defined [^(xxxi),^(xxxii),^(xxxiii),^(xxxiv)] Ligation activity (in addition to cleavage activity) makes ribozyme amenable to engineering through in vitro selection [^(xxxv)] Complete kinetic framework established for one ribozyme [^(xxxvi)]. Chemical modification investigation of important residues begun [^(xxxvii),^(xxxviii)]. Hepatitis Delta Virus (HDV) Ribozyme Size: ˜60 nucleotides. Trans cleavage of target RNAs demonstrated [^(xxxix)]. Binding sites and structural requirements not fully determined, although no sequences 5′ of cleavage site are required. Folded ribozyme contains a pseudoknot structure [^(xl)]. Reaction mechanism: attack by 2′-OH 5′ to the scissile bond to generate cleavage products with 2′,3′-cyclic phosphate and 5′-OH ends. Only 2 known members of this class. Found in human HDV. Circular form of HDV is active and shows increased nuclease stability [^(xli)]

[0229] TABLE II Reagent Equivalents Amount Wait Time* DNA Wait Time* 2′-O-methyl Wait Time*RNA A. 2.5 μmol Synthesis Cycle ABI 394 Instrument Phosphoramidites 6.5 163 μL 45 sec 2.5 min 7.5 min S-Ethyl Tetrazole 23.8 238 μL 45 sec 2.5 min 7.5 min Acetic Anhydride 100 233 μL  5 sec 5 sec 5 sec N-Methyl 186 233 μL  5 sec 5 sec 5 sec Imidazole TCA 176 2.3 mL 21 sec 21 sec 21 sec Iodine 11.2 1.7 mL 45 sec 45 sec 45 sec Beaucage 12.9 645 μL 100 sec  300 sec 300 sec Acetonitrile NA 6.67 mL NA NA NA B. 0.2 μmol Synthesis Cycle ABI 394 Instrument Phosphoramidites 15 31 μL 45 sec 233 sec 465 sec S-Ethyl Tetrazole 38.7 31 μL 45 sec 233 min 465 sec Acetic Anhydride 655 124 μL  5 sec 5 sec 5 sec N-Methyl 1245 124 μL  5 sec 5 sec 5 sec Imidazole TCA 700 732 μL 10 sec 10 sec 10 sec Iodine 20.6 244 μL 15 sec 15 sec 15 sec Beaucage 7.7 232 μL 100 sec  300 sec 300 sec Acetonitrile NA 2.64 mL NA NA NA C. 0.2 μmol Synthesis Cycle 96 well Instrument Equivalents: DNA/ Amount: DNA/2′-O- Wait Time* 2′-O- Wait Time* Reagent 2′-O-methyl/Ribo methyl/Ribo Wait Time* DNA methyl Ribo Phosphoramidites 22/33/66  40/60/120 μL  60 sec 180 sec 360 sec  S-Ethyl Tetrazole 70/105/210 40/60/120 μL  60 sec 180 min 360 sec  Acetic Anhydride 265/265/265  50/50/50 μL  10 sec 10 sec 10 sec N-Methyl 502/502/502  50/50/50 μL  10 sec 10 sec 10 sec Imidazole TCA 238/475/475  250/500/500 μL  15 sec 15 sec 15 sec Iodine 6.8/6.8/6.8    80/80/80 μL  30 sec 30 sec 30 sec Beaucage 34/51/51  80/120/120 100 sec 200 sec 200 sec  Acetonitrile NA 1150/1150/1150 μL NA NA NA

[0230] TABLE III Human PTGDR Hammerhead Ribozyme and Substrate Sequence Seq Seq Pos Substrate ID Hammerhead Ribozyme ID 12 UUCUGGCU A UUUUCCUC 1 GAGGAAAA CUGAUGAGGCCGUUAGGCCGAA AGCCAGAA 228 14 CUGGCUAU U UUCCUCCU 2 AGGAGGAA CUGAUGAGGCCGUUAGGCCGAA AUAGCCAG 229 15 UGGCUAUU U UCCUCCUG 3 CAGGAGGA CUGAUGAGGCCGUUAGGCCGAA AAUAGCCA 230 16 GGCUAUUU U CCUCCUGC 4 GCAGGAGG CUGAUGAGGCCGUUAGGCCGAA AAAUAGCC 231 17 GCUAUUUU C CUCCUGCC 5 GGCAGGAG CUGAUGAGGCCGUUAGGCCGAA AAAAUAGC 232 20 AUUUUCCU C CUGCCGUU 6 AACGGCAG CUGAUGAGGCCGUUAGGCCGAA AGGAAAAU 233 28 CCUGCCGU U CCGACUCG 7 CGAGUCGG CUGAUGAGGCCGUUAGGCCGAA ACGGCAGG 234 29 CUGCCGUU C CGACUCGG 8 CCGAGUCG CUGAUGAGGCCGUUAGGCCGAA AACGGCAG 235 35 UUCCGACU C GGCACCAG 9 CUGGUGCC CUGAUGAGGCCGUUAGGCCGAA AGUCGGAA 236 47 ACCAGAGU C UGUCUCUA 10 UAGAGACA CUGAUGAGGCCGUUAGGCCGAA ACUCUGGU 237 51 GAGUCUGU C UCUACUGA 11 UCAGUAGA CUGAUGAGGCCGUUAGGCCGAA ACAGACUC 238 53 GUCUGUCU C UACUGAGA 12 UCUCAGUA CUGAUGAGGCCGUUAGGCCGAA AGACAGAC 239 55 CUGUCUCU A CUGAGAAC 13 GUUCUCAG CUGAUGAGGCCGUUAGGCCGAA AGAGACAG 240 73 CAGCGCGU C AGGGCCGA 14 UCGGCCCU CUGAUGAGGCCGUUAGGCCGAA ACGCGCUG 241 85 GCCGAGCU C UUCACUGG 15 CCAGUGAA CUGAUGAGGCCGUUAGGCCGAA AGCUCGGC 242 87 CGAGCUCU U CACUGGCC 16 GGCCAGUG CUGAUGAGGCCGUUAGGCCGAA AGAGCUCG 243 88 GAGCUCUU C ACUGGCCU 17 AGGCCAGU CUGAUGAGGCCGUUAGGCCGAA AAGAGCUC 244 100 GGCCUGCU C CGCGCUCU 18 AGAGCGCG CUGAUGAGGCCGUUAGGCCGAA AGCAGGCC 245 107 UCCGCGCU C UUCAAUGC 19 GCAUUGAA CUGAUGAGGCCGUUAGGCCGAA AGCGCGGA 246 109 CGCGCUCU U CAAUGCCA 20 UGGCAUUG CUGAUGAGGCCGUUAGGCCGAA AGAGCGCG 247 110 GCGCUCUU C AAUGCCAG 21 CUGGCAUU CUGAUGAGGCCGUUAGGCCGAA AAGAGCGC 248 130 CAGGCGCU C ACCCUGCA 22 UGCAGGGU CUGAUGAGGCCGUUAGGCCGAA AGCGCCUG 249 145 CAGAGCGU C CCGCCUCU 23 AGAGGCGG CUGAUGAGGCCGUUAGGCCGAA ACGCUCUG 250 152 UCCCGCCU C UCAAAGAG 24 CUCUUUGA CUGAUGAGGCCGUUAGGCCGAA AGGCGGGA 251 154 CCGCCUCU C AAAGAGGG 25 CCCUCUUU CUGAUGAGGCCGUUAGGCCGAA AGAGGCGG 252 178 CCGCGAGU U UAGAUAGG 26 CCUAUCUA CUGAUGAGGCCGUUAGGCCGAA ACUCGCGG 253 179 CGCGAGUU U AGAUAGGA 27 UCCUAUCU CUGAUGAGGCCGUUAGGCCGAA AACUCGCG 254 180 GCGAGUUU A GAUAGGAG 28 CUCCUAUC CUGAUGAGGCCGUUAGGCCGAA AAACUCGC 255 184 GUUUAGAU A GGAGGUUC 29 GAACCUCC CUGAUGAGGCCGUUAGGCCGAA AUCUAAAC 256 191 UAGGAGGU U CCUGCCGU 30 ACGGCAGG CUGAUGAGGCCGUUAGGCCGAA ACCUCCUA 257 192 AGGAGGUU C CUGCCGUG 31 CACGGCAG CUGAUGAGGCCGUUAGGCCGAA AACCUCCU 258 220 GCCGCCCU C GGAGCUUU 32 AAAGCUCC CUGAUGAGGCCGUUAGGCCGAA AGGGCGGC 259 227 UCGGAGCU U UUUCUGUG 33 CACAGAAA CUGAUGAGGCCGUUAGGCCGAA AGCUCCGA 260 228 CGGAGCUU U UUCUGUGG 34 CCACAGAA CUGAUGAGGCCGUUAGGCCGAA AAGCUCCG 261 229 GGAGCUUU U UCUGUGGC 35 GCCACAGA CUGAUGAGGCCGUUAGGCCGAA AAAGCUCC 262 230 GAGCUUUU U CUGUGGCG 36 CGCCACAG CUGAUGAGGCCGUUAGGCCGAA AAAAGCUC 263 231 AGCUUUUU C UGUGGCGC 37 GCGCCACA CUGAUGAGGCCGUUAGGCCGAA AAAAAGCU 264 244 GCGCAGCU U CUCCGCCC 38 GGGCGGAG CUGAUGAGGCCGUUAGGCCGAA AGCUGCGC 265 245 CGCAGCUU C UCCGCCCG 39 CGGGCGGA CUGAUGAGGCCGUUAGGCCGAA AAGCUGCG 266 247 CAGCUUCU C CGCCCGAG 40 CUCGGGCG CUGAUGAGGCCGUUAGGCCGAA AGAAGCUG 267 280 CGGGGGCU C CUUAGCAC 41 GUGCUAAG CUGAUGAGGCCGUUAGGCCGAA AGCCCCCG 268 283 GGGCUCCU U AGCACCCG 42 CGGGUGCU CUGAUGAGGCCGUUAGGCCGAA AGGAGCCC 269 284 GGCUCCUU A GCACCCGG 43 CCGGGUGC CUGAUGAGGCCGUUAGGCCGAA AAGGAGCC 270 306 GGGGCCCU C GCCCUUCC 44 GGAAGGGC CUGAUGAGGCCGUUAGGCCGAA AGGGCCCC 271 312 CUCGCCCU U CCGCAGCC 45 GGCUGCGG CUGAUGAGGCCGUUAGGCCGAA AGGGCGAG 272 313 UCGCCCUU C CGCAGCCU 46 AGGCUGCG CUGAUGAGGCCGUUAGGCCGAA AAGGGCGA 273 322 CGCAGCCU U CACUCCAG 47 CUGGAGUG CUGAUGAGGCCGUUAGGCCGAA AGGCUGCG 274 323 GCAGCCUU C ACUCCAGC 48 GCUGGAGU CUGAUGAGGCCGUUAGGCCGAA AAGGCUGC 275 327 CCUUCACU C CAGCCCUC 49 GAGGGCUG CUGAUGAGGCCGUUAGGCCCAA AGUGAAGG 276 335 CCAGCCCU C UGCUCCCG 50 CGUGAGCA CUGAUGAGGCCGUUACGCCGAA AGGGCUGG 277 340 CCUCUGCU C CCGCACGC 51 GCGUGCGG CUGAUGAGGCCGUUAGGCCGAA AGCAGAGG 278 357 CAUGAAGU C GCCGUUCU 52 AGAACGGC CUGAUGAGGCCGUUAGGCCGAA ACUUCAUG 279 363 GUCGCCGU U CUACCGCU 53 ACCCGUAG CUGAUGAGGCCGUUAGGCCGAA ACGGCGAC 280 364 UCGCCGUU C UACCGCUG 54 CAGCGGUA CUGAUGAGGCCGUUAGGCCGAA AACGCCGA 281 366 GCCGUUCU A CCGCUGCC 55 GGCAGCGG CUGAUGAGGCCGUUAGGCCGAA AGAACGGC 282 387 CACCACCU C UCUCGAAA 56 UUUCCACA CUGAUGAGGCCGUUAGGCCGAA AGGUGGUG 283 405 AGGCAACU C GGCGGUGA 57 UCACCGCC CUGAUGAGGCCGUUAGGCCGAA AGUUGCCU 284 427 GCGGUGCU C UUCAGCAC 58 GUGCUGAA CUGAUGAGGCCGUUAGGCCGAA AGCACCCC 285 429 GGUGCUCU U CAGCACCG 59 CGGUGCUG CUGAUGAGGCCUUUAGGCCGAA AGAGCACC 286 430 GUGCUCUU C AGCACCGG 60 CCGGUGCU CUGAUGAGGCCGUUAGGCCGAA AAGAGCAC 287 442 ACCGGCCU C CUGGGCAA 61 UUGCCCAG CUGAUGAGGCCGUUAGGCCGAA AGGCCGGU 288 480 GGCCCGCU C GGGGCUGG 62 CCAGCCCC CUGAUGAGGCCGUUAGGCCGAA AGCGCGCC 289 498 GUGGUGCU C GCGGCGUC 63 GACGCCGC CUGAUGAGGCCGUUAGGCCGAA AGCACCAC 290 506 CGCGGCGU C CACUGCGC 64 GCGCAGUG CUGAUGAGGCCGUUAGGCCCAA ACGCCGCG 291 525 GCUGCCCU C GGUCUUCU 65 AGAAGACC CUGAUGAGGCCGUUAGGCCGAA AGGGCAGC 292 529 CCCUCGGU C UUCUACAU 66 AUGUAGAA CUGAUGAGGCCGUUAGGCCGAA ACCGAGGG 293 531 CUCGGUCU U CUACAUGC 67 GCAUGUAG CUGAUGAGGCCCUUACGCCGAA AGACCGAG 294 532 UCGGUCUU C UACAUGCU 68 AGCAUGUA CUGAUGAGGCCGUUAGGCCGAA AAGACCGA 295 534 GGUCUUCU A CAUGCUGG 69 CCAGCAUG CUGAUGAGGCCGUUAGCCCGAA AGAAGACC 296 559 CUGACGGU C ACCGACUU 70 AAGUCGGU CUGAUGAGGCCGUUAGGCCGAA ACCGUCAG 297 567 CACCGACU U GCUGGGCA 71 UCCCCAGC CUGAUCAGCCCGUUAGGCCCAA AGUCGGUG 298 583 AAGUGCCU C CUAAGCCC 72 GGGCUUAG CUGAUGAGGCCGUUAGGCCGAA AGGCACUU 299 586 UGCCUCCU A AGCCCGGU 73 ACCGGGCU CUGAUGAGGCCCUUAGGCCGAA ACGAGGCA 300 609 GGCUGCCU A CGCUCAGA 74 UCUGAGCG CUGAUGAGGCCGUUAGGCCGAA AGGCAGCC 301 614 CCUACCCU C AGAACCGG 75 CCGGUUCU CUGAUGAGGCCGUUAGGCCGAA AGCGUAGG 302 626 ACCGGAGU C UGCGGGUG 76 CACCCCCA CUGAUGAGGCCCUUAGGCCGAA ACUCCCCU 303 637 CGCGUGCU U CCGCCCGC 77 GCGGGCGC CUGAUGAGGCCGUUAGGCCGAA AGCACCCG 304 648 GCCCGCAU U GGACAACU 78 AGUUGUCC CUGAUGAGGCCGUUAGGCCGAA AUGCGGGC 305 657 CGACAACU C GUUGUGCC 79 GGCACAAC CUGAUGAGGCCGUUAGGCCGAA AGUUGUCC 306 660 CAACUCGU U GUGCCAAG 80 CUUGGCAC CUGAUGAGGCCGUUAGGCCGAA ACGAGUUG 307 672 CCAAGCCU U CGCCUUCU 81 AGAAGGCG CUGAUGAGGCCGUUAGGCCGAA AGGCUUGG 308 673 CAAGCCUU C GCCUUCUU 82 AAGAAGGC CUGAUGAGGCCGUUAGGCCGAA AAGGCUUG 309 678 CUUCGCCU U CUUCAUGU 83 ACAUGAAG CUGAUGAGGCCGUUAGGCCGAA AGGCGAAG 310 679 UUCGCCUU C UUCAUGUC 84 GACAUGAA CUGAUGAGGCCGUUAGGCCGAA AAGGCGAA 311 681 CGCCUUCU U CAUGUCCU 85 AGGACAUC CUGAUGACCCCGUUAGGCCGAA ACAAGGCC 312 682 GCCUUCUU C AUGUCCUU 86 AAGGACAU CUGAUGAGGCCGUUAGGCCGAA AAGAAGGC 313 687 CUUCAUGU C CUUCUUUG 87 CAAAGAAG CUGAUGAGGCCGUUAGGCCGAA ACAUGAAG 314 690 CAUGUCCU U CUUUGGGC 88 GCCCAAAG CUGAUGAGGCCGUUAGGCCGAA AGGACAUG 315 691 AUGUCCUU C UUUGGGCU 89 AGCCCAAA CUGAUGAGGCCGUUAGGCCGAA AAGGACAU 316 693 GUCCUUCU U UGGGCUCU 90 AGAGCCCA CUGAUGAGGCCGUUAGGCCGAA AGAAGGAC 317 694 UCCUUCUU U GGGCUCUC 91 GAGAGCCC CUGAUGAGGCCGUUAGGCCGAA AAGAAGGA 318 700 UUUGGGCU C UCCUCGAC 92 GUCGAGGA CUGAUGAGGCCGUUAGGCCGAA AGCCCAAA 319 702 UGGGCUCU C CUCGACAC 93 GUGUCGAG CUGAUGAGGCCGUUAGGCCGAA AGAGCCCA 320 705 GCUCUCCU C GACACUGC 94 GCAGUGUC CUGAUGAGGCCGUUAGGCCGAA AGGAGAGC 321 718 CUGCAACU C CUGGCCAU 95 AUGGCCAG CUGAUGAGGCCGUUAGGCCGAA AGUUGCAG 322 745 UGCUGGCU C UCCCUAGG 96 CCUAGGGA CUGAUGAGGCCGUUAGGCCGAA AGCCAGCA 323 747 CUGGCUCU C CCUAGGGC 97 GCCCUAGG CUGAUGAGGCCGUUAGGCCGAA AGAGCCAG 324 751 CUCUCCCU A GGGCACCC 98 GGGUGCCC CUGAUGAGGCCGUUAGGCCGAA AGGGAGAG 325 761 GGCACCCU U UCUUCUAC 99 GUAGAAGA CUGAUGAGGCCGUUAGGCCGAA AGGGUGCC 326 762 GCACCCUU U CUUCUACC 100 GGUAGAAG CUGAUGAGGCCGUUAGGCCGAA AAGGGUGC 327 763 CACCCUUU C UUCUACCC 101 CGGUAGAA CUGAUGAGCCCGUUAGGCCGAA AAAGGGUG 328 765 CCCUUUCU U CUACCGAC 102 GUCGGUAG CUGAUGAGGCCGUUAGGCCGAA AGAAAGGG 329 766 CCUUUCUU C UACCGACG 103 CGUCGGUA CUGAUGAGGCCGUUAGGCCGAA AAGAAAGG 330 768 UUUCUUCU A CCGACGGC 104 GCCGUCGG CUCAUGAGGCCUUUAGGCCGAA AGAAGAAA 331 781 CGCCACAU C ACCCUGCG 105 CGCAGGGU CUGAUGAGGCCGUUAGGCCGAA AUGUGCCG 332 825 GAGCGCCU U CUCCCUGG 106 CCAGGGAG CUGAUCAGCCCGUUAGGCCGAA AGGCGCUC 333 826 AGCGCCUU C UCCCUGGC 107 GCCAGGGA CUGAUGAGGCCGUUAGGCCGAA AAGGCGCU 334 828 CGCCUUCU C CCUGGCUU 108 AAGCCACG CUGAUGAGGCCGUUAGGCCGAA AGAAGGCG 335 836 CCCUGGCU U UCUGCGCG 109 CGCGCAGA CUGAUGAGGCCGUUAGGCCGAA AUCCAGGG 336 837 CCUGGCUU U CUGCGCGC 110 GCGCGCAG CUGAUCACGCCGUUAGGCCGAA AAGCCAGG 337 838 CUGGCUUU C UGCGCGCU 111 AGCGCGCA CUGAUGAGGCCGUUAGGCCGAA AAAGCCAG 338 847 UGCGCGCU A CCUUUCAU 112 AUGAAAGC CUCAUGAGGCCGUUACGCCGAA AGCGCGCA 339 851 CGCUACCU U UCAUCGCC 113 GCCCAUGA CUGAUGAGGCCGUUAGGCCGAA AGGUAGCG 340 852 GCUACCUU U CAUGGGCU 114 AGCCCAUG CUGAUGAGGCCGUUAGGCCGAA AAGGUAGC 341 853 CUACCUUU C AUGGGCUU 115 AAGCCCAU CUGAUGAGGCCGUUAGGCCGAA AAAGGUAG 342 861 CAUGGGCU U CGGGAAGU 116 ACUUCCCG CUGAUGAGGCCGUUAGGCCGAA AGCCCAUG 343 862 AUGGGCUU C GGGAAGUU 117 AACUUCCC CUGAUGAGGCCGUUAGGCCGAA AAGCCCAU 344 870 CGGGAAGU U CGUGCAGU 118 ACUGCACG CUGAUGAGGCCGUUAGGCCGAA ACUUCCCG 345 871 GGGAAGUU C GUGCAGUA 119 UACUGCAC CUGAUGAGGCCGUUAGGCCGAA AACUUCCC 346 879 CGUGCAGU A CUGCCCCG 120 CGGGGCAG CUGAUGAGGCCGUUAGGCCGAA ACUGCACG 347 900 CUGGUGCU U UAUCCAGA 121 UCUGGAUA CUGAUGAGGCCGUUAGGCCGAA AGCACCAG 348 901 UGGUGCUU U AUCCAGAU 122 AUCUGGAU CUGAUGAGGCCGUUAGGCCGAA AAGCACCA 349 902 GGUGCUUU A UCCAGAUG 123 CAUCUGGA CUGAUGAGGCCGUUAGGCCGAA AAAGCACC 350 904 UGCUUUAU C CAGAUGGU 124 ACCAUCUG CUGAUGAGGCCGUUAGGCCGAA AUAAAGCA 351 913 CAGAUGGU C CACGAGGA 125 UCCUCGUG CUGAUGAGGCCGUUAGGCCGAA ACCAUCUG 352 927 GGAGGGCU C GCUGUCGG 126 CCGACAGC CUGAUGAGGCCGUUAGGCCGAA AGCCCUCC 353 933 CUCGCUGU C GGUGCUGG 127 CCAGCACC CUGAUGAGGCCGUUAGGCCGAA ACAGCGAG 354 945 GCUGGGGU A CUCUGUGC 128 GCACAGAG CUGAUGAGGCCGUUAGGCCGAA ACCCCAGC 355 948 GGGGUACU C UGUGCUCU 129 AGAGCACA CUGAUGAGGCCGUUAGGCCGAA AGUACCCC 356 955 UCUGUGCU C UACUCCAG 130 CUGGAGUA CUGAUGAGGCCGUUAGGCCGAA AGCACAGA 357 957 UGUGCUCU A CUCCAGCC 131 GGCUGGAG CUGAUGAGGCCGUUAGGCCGAA AGAGCACA 358 960 GCUCUACU C CAGCCUCA 132 UGAGGCUG CUGAUGAGGCCGUUAGGCCGAA AGUAGAGC 359 967 UCCAGCCU C AUGGCGCU 133 AGCGCCAU CUGAUGAGGCCGUUAGGCCGAA AGGCUGGA 360 982 CUGCUGGU C CUCGCCAC 134 GUGGOGAG CUGAUGAGGCCGUUAGGCCGAA ACCAGCAG 361 985 CUGGUCCU C GCCACCGU 135 ACGGUGGC CUGAUGAGGCCGUUAGGCCGAA AGGACCAG 362 1006 UGCAACCU C GGCGCCAU 136 AUGGCGCC CUGAUGAGGCCGUUAGGCCGAA AGGUUGCA 363 1024 CGCAACCU C UAUGCGAU 137 AUCGCAUA CUGAUGAGGCCGUUAGGCCGAA AGGUUGCG 364 1026 CAACCUCU A UGCGAUGC 138 GCAUCGCA CUGAUGAGGCCGUUAGGCCGAA AGAGGUUG 365 1062 CCCGCGCU C CUCCACCA 139 UGGUGCAG CUGAUGAGGCCGUUAGGCCGAA AGCGCGGG 366 1110 GGAAGCGU C CCCUCAGC 140 GCUGAGGG CUGAUGAGGCCGUUAGGCCGAA ACGCUUCC 367 1115 CGUCCCCU C AGCCCCUG 141 CAGGGGCU CUGAUGAGGCCGUUAGGCCGAA AGGGGACG 368 1136 AGCUGGAU C ACCUCCUG 142 CAGGAGGU CUGAUGAGGCCGUUAGGCCGAA AUCCAGCU 369 1141 GAUCACCU C CUGCUGCU 143 AGCAGCAG CUGAUGAGGCCGUUAGGCCGAA AGGUGAUC 370 1168 ACCGUGCU C UUCACUAU 144 AUAGUGAA CUGAUGAGGCCGUUAGGCCGAA AGCACGGU 371 1170 CGUGCUCU U CACUAUGU 145 ACAUAGUG CUGAUGAGGCCGUUAGGCCGAA AGAGCACG 372 1171 GUGCUCUU C ACUAUGUG 146 CACAUAGU CUGAUGAGGCCGUUAGGCCGAA AAGAGCAC 373 1175 UCUUCACU A UGUGUUCU 147 AGAACACA CUGAUGAGGCCGUUAGGCCGAA AGUGAAGA 374 1181 CUAUGUGU U CUCUGCCC 148 GGGCAGAG CUGAUGAGGCCGUUAGGCCGAA ACACAUAG 375 1182 UAUGUGUU C UCUGCCCG 149 CGGGCAGA CUGAUGAGGCCGUUAGGCCGAA AACACAUA 376 1184 UGUGUUCU C UGCCCGUA 150 UACGGGCA CUGAUGAGCCCGUUAGGCCGAA AGAACACA 377 1192 CUGCCCGU A AUUUAUCG 151 CGAUAAAU CUGAUGAGGCCGUUAGGCCGAA ACGGGCAC 378 1195 CCCGUAAU U UAUCGCGC 152 GCGCGAUA CUGAUGAGGCCGUUAGGCCGAA AUUACGGG 379 1196 CCGUAAUU U AUCGCGCU 153 AGCGCGAU CUCAUGAGGCCGUUAGGCCGAA AAUUACGG 380 1197 CGUAAUUU A UCGCUCUU 154 AAGCGCGA CUGAUGAGGCCGUUAGGCCGAA AAAUUACG 381 1199 UAAUUUAU C GCGCUUAC 155 GUAAGCGC CUGAUGAGGCCGUUAGGCCGAA AUAAAUUA 382 1205 AUCGCGCU U ACUAUGGA 156 UCCAUAGU CUGAUGAGGCCGUUAGGCCGAA AGCGCGAU 383 1206 UCGCGCUU A CUAUGGAG 157 CUCCAUAG CUGAUGAGGCCGUUAGGCCGAA AAGCGCGA 384 1209 CGCUUACU A UGGAGCAU 158 AUGCUCCA CUGAUGAGGCCGUUAGGCCGAA AGUAAGCG 385 1218 UGGAGCAU U UAAGGAUG 159 CAUCCUUA CUGAUGAGGCCCUUAGGCCGAA AUGCUCCA 386 1219 GGAGCAUU U AAGGAUGU 160 ACAUCCUU CUGAUGAGGCCGUUAGGCCGAA AAUGCUCC 387 1220 GAGCAUUU A AGGAUGUC 161 GACAUCCU CUGAUGAGGCCGUUAGGCCGAA AAAUGCUC 388 1228 AAGGAUGU C AAGGAGAA 162 UUCUCCUU CUGAUGAGGCCGUUAGGCCGAA ACAUCCUC 389 1248 CAGGACCU C UGAAGAAG 163 CUUCUUCA CUGAUGAGGCCGUUAGGCCGAA AGGUCCUG 390 1267 GAAGACCU C CGAGCCUU 164 AAGGCUCG CUGAUGAGGCCGUUAGGCCGAA AGGUCUUC 391 1275 CCGAGCCU U GCGAUUUC 165 GAAAUCGC CUGAUGAGGCCGUUAGGCCGAA AGGCUCGG 392 1281 CUUGCGAU U UCUAUCUG 166 CAGAUAGA CUGAUGAGGCCGUUAGGCCGAA AUCGCAAG 393 1282 UUGCGAUU U CUAUCUGU 167 ACAGAUAG CUGAUGAGGCCGUUAGGCCGAA AAUCGCAA 394 1283 UGCGAUUU C UAUCUGUG 168 CACAGAUA CUGAUGAGGCCGUUAGGCCGAA AAAUCGCA 395 1285 CGAUUUCU A UCUGUGAU 169 AUCACAGA CUGAUGAGGCCGUUAGGCCGAA AGAAAUCG 396 1287 AUUUCUAU C UGUGAUUU 170 AAAUCACA CUGAUGAGGCCGUUAGGCCGAA AUAGAAAU 397 1294 UCUGUGAU U UCAAUUGU 171 ACAAUUGA CUGAUGAGGCCGUUAGGCCGAA AUCACAGA 398 1295 CUGUGAUU U CAAUUGUG 172 CACAAUUG CUGAUGAGGCCGUUAGGCCGAA AAUCACAG 399 1296 UGUGAUUU C AAUUGUGG 173C CACAAUU CUGAUGAGGCCGUUAGGCCGAA AAAUCACA 400 1300 AUUUCAAU U GUGGACCC 174 GGGUCCAC CUGAUGAGGCCGUUAGGCCGAA AUUGAAAU 401 1310 UGGACCCU U GGAUUUUU 175 AAAAAUCC CUGAUGAGGCCGUUAGGCCGAA AGGGUCCA 402 1315 CCUUGGAU U UUUAUCAU 176 AUGAUAAA CUGAUGAGGCCGUUAGGCCGAA AUCCAAGG 403 1316 CUUGGAUU U UUAUCAUU 177 AAUGAUAA CUGAUGAGGCCGUUAGGCCGAA AAUCCAAG 404 1317 UUGGAUUU U UAUCAUUU 178 AAAUGAUA CUGAUGAGGCCGUUAGGCCGAA AAAUCCAA 405 1318 UGGAUUUU U AUCAUUUU 179 AAAAUGAU CUGAUGAGGCCGUUAGGCCGAA AAAAUCCA 406 1319 GGAUUUUU A UCAUUUUC 180 GAAAAUGA CUGAUGAGGCCGUUAGGCCGAA AAAAAUCC 407 1321 AUUUUUAU C AUUUUCAG 181 CUGAAAAU CUGAUGAGGCCGUUAGGCCGAA AUAAAAAU 408 1324 UUUAUCAU U UUCAGAUC 182 GAUCUGAA CUGAUGAGGCCGUUAGGCCGAA AUGAUAAA 409 1325 UUAUCAUU U UCAGAUCU 183 AGAUCUGA CUGAUGAGGCCGUUAGGCCGAA AAUGAUAA 410 1326 UAUCAUUU U CAGAUCUC 184 GAGAUCUG CUGAUGAGGCCGUUAGGCCGAA AAAUGAUA 411 1327 AUCAUUUU C AGAUCUCC 185 GGAGAUCU CUGAUGAGGCCGUUAGGCCGAA AAAAUGAU 412 1332 UUUCAGAU C UCCAGUAU 186 AUACUGGA CUGAUGAGGCCGUUAGGCCGAA AUCUGAAA 413 1334 UCAGAUCU C CAGUAUUU 187 AAAUACUG CUGAUGAGGCCGUUAGGCCGAA AGAUCUGA 414 1339 UCUCCAGU A UUUCGGAU 188 AUCCGAAA CUGAUGAGGCCGUUAGGCCGAA ACUGGAGA 415 1341 UCCAGUAU U UCGGAUAU 189 AUAUCCGA CUGAUGAGGCCGUUAGGCCGAA AUACUGGA 416 1342 CCAGUAUU U CGGAUAUU 190 AAUAUCCG CUGAUGAGGCCGUUAGGCCGAA AAUACUGG 417 1343 CAGUAUUU C GGAUAUUU 191 AAAUAUCC CUGAUGAGGCCGUUAGGCCGAA AAAUACUG 418 1348 UUUCGGAU A UUUUUUCA 192 UGAAAAAA CUGAUGAGGCCGUUAGGCCGAA AUCCGAAA 419 1350 UCGGAUAU U UUUUCACA 193 UGUGAAAA CUGAUGAGGCCGUUAGGCCGAA AUAUCCGA 420 1351 CGGAUAUU U UUUCACAA 194 UUGUGAAA CUGAUGAGGCCGUUAGGCCGAA AAUAUCCG 421 1352 GGAUAUUU U UUCACAAG 195 CUUGUGAA CUGAUGAGGCCGUUAGGCCGAA AAAUAUCC 422 1353 GAUAUUUU U UCACAAGA 196 UCUUGUGA CUGAUGAGGCCGUUAGGCCGAA AAAAUAUC 423 1354 AUAUUUUU U CACAAGAU 197 AUCUUGUG CUGAUGAGGCCGUUAGGCCGAA AAAAAUAU 424 1355 UAUUUUUU C ACAAGAUU 198 AAUCUUGU CUGAUGAGGCCGUUAGGCCGAA AAAAAAUA 425 1363 CACAAGAU U UUCAUUAG 199 CUAAUGAA CUGAUGAGGCCGUUAGGCCGAA AUCUUGUG 426 1364 ACAAGAUU U UCAUUAGA 200 UCUAAUGA CUGAUGAGGCCGUUAGGCCGAA AAUCUUGU 427 1365 CAAGAUUU U CAUUAGAC 201 GUCUAAUG CUGAUGAGGCCGUUAGGCCGAA AAAUCUUG 428 1366 AAGAUUUU C AUUAGACC 202 CGUCUAAU CUGAUGAGGCCGUUACGCCGAA AAAAUCUU 429 1369 AUUUUCAU U AGACCUCU 203 AGAGGUCU CUGAUGAGGCCGUUAGGCCGAA AUGAAAAU 430 1370 UUUUCAUU A GACCUCUU 204 AAGAGGUC CUGAUGAGGCCGUUAGGCCGAA AAUGAAAA 431 1376 UUAGACCU C UUAGGUAC 205 GUACCUAA CUGAUGAGGCCGUUAGGCCGAA AGGUCUAA 432 1378 AGACCUCU U AGGUACAG 206 CUGUACCU CUGAUGAGGCCGUUAGGCCGAA AGAGGUCU 433 1379 GACCUCUU A GGUACAGG 207 CCUGUACC CUGAUGAGGCCGUUAGGCCGAA AAGAGGUC 434 1383 UCUUAGGU A CAGGAGCC 208 GGCUCCUG CUGAUGAGGCCGUUAGGCCGAA ACCUAAGA 435 1403 GCAGCAAU U CCACUAAC 209 GUUAGUGG CUGAUGAGGCCGUUAGGCCGAA AUUGCUGC 436 1404 CAGCAAUU C CACUAACA 210 UGUUAGUG CUGAUGAGGCCGUUAGGCCGAA AAUUGCUG 437 1409 AUUCCACU A ACAUGGAA 211 UUCCAUGU CUGAUGAGGCCGUUAGGCCGAA AGUGGAAU 438 1419 CAUGGAAU C CAGUCUGU 212 ACAGACUG CUGAUGAGGCCGUUAGGCCGAA AUUCCAUG 439 1424 AAUCCAGU C UGUGACAG 213 CUGUCACA CUGAUGAGGCCGUUAGGCCGAA ACUGGAUU 440 1436 GACAGUGU U UUUCACUC 214 GAGUGAAA CUGAUGAGGCCGUUAGGCCGAA ACACUGUC 441 1437 ACAGUGUU U UUCACUCU 215 AGAGUGAA CUGAUGAGGCCGUUAGGCCGAA AACACUGU 442 1438 CAGUGUUU U UCACUCUG 216 CAGAGUGA CUGAUGAGGCCGUUAGGCCGAA AAACACUG 443 1439 AGUGUUUU U CACUCUGU 217 ACAGAGUG CUGAUGAGGCCGUUAGGCCGAA AAAACACU 444 1440 GUGUUUUU C ACUCUGUG 218 CACAGAGU CUGAUGAGGCCGUUAGGCCGAA AAAAACAC 445 1444 UUUUCACU C UGUGGUAA 219 UUACCACA CUGAUGAGGCCGUUAGGCCGAA AGUGAAAA 446 1451 UCUGUGGU A AGCUGAGG 220 CCUCAGCU CUGAUGAGGCCGUUAGGCCGAA ACCACAGA 447 1463 UGAGGAAU A UGUCACAU 221 AUGUGACA CUGAUGAGGCCGUUAGGCCGAA AUUCCUCA 448 1467 GAAUAUGU C ACAUUUUC 222 GAAAAUGU CUGAUGAGGCCGUUAGGCCGAA ACAUAUUC 449 1472 UGUCACAU U UUCAGUCA 223 UGACUGAA CUGAUGAGGCCGUUAGGCCGAA AUGUGACA 450 1473 GUCACAUU U UCAGUCAA 224 UUGACUGA CUGAUGAGGCCGUUAGGCCGAA AAUGUGAC 451 1474 UCACAUUU U CAGUCAAA 225 UUUGACUG CUGAUGAGGCCGUUAGGCCGAA AAAUGUGA 452 1475 CACAUUUU C AGUCAAAG 226 CUUUGACU CUGAUGAGGCCGUUAGGCCGAA AAAAUGUG 453 1479 UUUUCAGU C AAAGAACC 227 GGUUCUUU CUGAUGAGGCCGUUAGGCCGAA ACUGAAAA 454

[0231] TABLE IV Human PTGDR Inozyme and Substrate Sequence Seq Seq Pos Substrate ID Inozyme ID 11 AUUCUGGC U AUUUUCCU 455 AGGAAAAU CUGAUGAGGCCGUUAGGCCGAA ICCAGAAU 831 18 CUAUUUUC C UCCUGCCG 456 CGGCAGGA CUGAUGAGGCCGUUAGGCCGAA IAAAAUAG 832 19 UAUUUUCC U CCUGCCGU 457 ACGGCAGG CUGAUGAGGCCGUUAGGCCGAA IGAAAAUA 833 21 UUUUCCUC C UGCCGUUC 458 GAACGGCA CUGAUGAGGCCGUUAGGCCGAA IAGGAAAA 834 22 UUUCCUCC U GCCGUUCC 459 GGAACGGC CUGAUGAGGCCGUUAGGCCGAA IGAGGAAA 835 25 CCUCCUGC C GUUCCGAC 460 GUCGGAAC CUGAUCACGCCGUUAGGCCCAA ICAGGAGG 836 30 UGCCGUUC C GACUCCUC 461 GCCGAGUC CUGAUGAGGCCGUUAGGCCGAA IAACGGCA 837 34 GUUCCGAC U CGGCACCA 462 UGGUGCCG CUGAUGAGGCCGUUAGGCCGAA IUCGGAAC 838 39 GACUCGGC A CCAGAGUC 463 GACUCUGG CUGAUGAGGCCGUUAGGCCGAA ICCGAGUC 839 41 CUCGGCAC C AGAGUCUG 464 CAGACUCU CUGAUGAGGCCGUUAGGCCGAA IUGCCGAG 840 42 UCGGCACC A GAGUCUGU 465 ACAGACUC CUGAUGAGGCCGUUAGGCCGAA IGUGCCGA 841 48 CCAGAGUC U GUCUCUAC 466 GUAGAGAC CUGAUGAGGCCGUUAGGCCGAA IACUCUGG 842 52 AGUCUGUC U CUACUGAG 467 CUCAGUAG CUGAUGAGGCCGUUAGGCCGAA IACAGACU 843 54 UCUGUCUC U ACUGAGAA 468 UUCUCAGU CUGAUGAGGCCGUUAGGCCGAA IAGACAGA 844 57 GUCUCUAC U GAGAACGC 469 GCGUUCUC CUGAUGAGGCCGUUAGGCCGAA IUAGAGAC 845 66 GAGAACGC A GCGCGUCA 470 UGACGCGC CUGAUGAGGCCGUUAGGCCGAA ICGUUCUC 846 74 AGCGCGUC A GGGCCGAG 471 CUCGGCCC CUGAUGAGGCCGUUAGGCCGAA IACGCGCU 847 79 GUCAGGGC C GAGCUCUU 472 AAGAGCUC CUGAUGAGGCCGUUAGGCCGAA ICCCUGAC 848 84 GGCCGAGC U CUUCACUG 473 CAGUGAAG CUGAUGAGGCCGUUAGGCCGAA ICUCGGCC 849 86 CCGAGCUC U UCACUGGC 474 GCCAGUGA CUGAUGAGGCCGUUAGGCCGAA IAGCUCGG 850 89 AGCUCUUC A CUGGCCUG 475 CAGGCCAG CUGAUGAGGCCGUUAGGCCGAA IAAGAGCU 851 91 CUCUUCAC U GGCCUGCU 476 AGCAGGCC CUGAUGAGGCCGUUAGGCCGAA IUGAAGAG 852 95 UCACUGGC C UGCUCCGC 477 GCGGAGCA CUGAUGAGGCCGUUAGGCGGAA ICCAGUGA 853 96 CACUGGCC U GCUCCGCG 478 CGCGGAGC CUGAUGAGGCCGUUAGGCCGAA IGCCAGUG 854 99 UGGCCUGC U CCGCGCUC 479 GAGCGCGG CUGAUGAGGCCGUUAGGCCGAA ICAGGCCA 855 101 GCCUGCUC C GCGCUCUU 480 AAGAGCGC CUGAUGAGGCCGUUAGGCCGAA IAGCAGGC 856 106 CUCCGCGC U CUUCAAUG 481 CAUUGAAG CUGAUGAGGCCGUUAGGCCGAA ICGCGGAG 857 108 CCGCGCUC U UCAAUGCC 482 GGCAUUGA CUGAUGAGGCCGUUAGGCCGAA IAGCGCGG 858 111 CGCUCUUC A AUGCCAGC 483 GCUGGCAU CUGAUGAGGCCGUUAGGCCGAA IAAGAGCG 859 116 UUCAAUGC C AGCGCCAG 484 CUGGCGCU CUGAUGAGGCCGUUAGGCCGAA ICAUUGAA 860 117 UCAAUGCC A GCGCCAGG 485 CCUGGCGC CUGAUGAGGCCGUUAGGCCGAA IGCAUUGA 861 122 GCCAGCGC C AGGCGCUC 486 GAGCGCCU CUGAUGAGGCCGUUAGGCCGAA ICGCUGGC 862 123 CCAGCGCC A GGCGCUCA 487 UGAGCGCC CUGAUGAGGCCGUUAGGCCGAA IGCGCUGG 863 129 CCAGGCGC U CACCCUGC 488 GCAGGGUG CUGAUGAGGCCGUUAGGCCGAA ICGCCUGG 864 131 AGGCGCUC A CCCUGCAG 489 CUGCAGGG CUGAUGAGGCCGUUAGGCCGAA IAGCGCCU 865 133 GCGCUCAC C CUGCAGAG 490 CUCUGCAG CUGAUGAGGCCGUUAGGCCGAA IUGAGCGC 866 134 CGCUCACC C UGCAGAGC 491 GCUCUGCA CUGAUGAGGCCGUUAGGCCGAA IGUGAGCG 867 135 GCUCACCC U GCAGAGCG 492 CGCUCUGC CUGAUGAGGCCGUUAGGCCGAA IGGUGAGC 868 138 CACCCUGC A GAGCGUCC 493 GGACGCUC CUGAUGAGGCCGUUAGGCCGAA ICAGGGUG 869 146 AGAGCGUC C CGCCUCUC 494 GAGAGGCG CUGAUGAGGCCGUUAGGCCGAA IACGCUCU 870 147 GAGCGUCC C GCCUCUCA 495 UGAGAGGC CUGAUGAGGCCGUUAGGCCGAA IGACGCUC 871 150 CGUCCCGC C UCUCAAAG 496 CUUUGAGA CUGAUGAGGCCGUUAGGCCGAA ICGGGACG 872 151 GUCCCGCC U CUCAAAGA 497 UCUUUGAG CUGAUGAGGCCGUUAGGCCGAA IGCGGGAC 873 153 CCCGCCUC U CAAAGAGG 498 CCUCUUUG CUGAUGAGGCCGUUAGGCCGAA IAGGCGGG 874 155 CGCCUCUC A AAGAGGGG 499 CCCCUCUU CUGAUGAGGCCGUUAGGCCGAA IAGAGGCG 875 170 GGUGUGAC C CGCGAGUU 500 AACUCGCG CUGAUGAGGCCGUUAGGCCGAA IUCACACC 876 171 GUGUGACC C GCGAGUUU 501 AAACUCGC CUGAUGAGGCCGUUAGGCCGAA IGUCACAC 877 193 GGAGGUUC C UGCCCUCG 502 CCACGGCA CUCAUGAGGCCGUUAGGCCGAA IAACCUCC 878 194 GACGUUCC U GCCGUGGG 503 CCCACGGC CUGAUGAGGCCGUUAGGCCGAA IGAACCUC 879 197 GUUCCUGC C GUGGGGAA 504 UUCCCCAC CUGAUGAGGCCGUUAGGCCGAA ICAGGAAC 880 207 UGCGGAAC A CCCCGCCG 505 CCGCGGGG CUGAUGAGGCCGUUAGGCCGAA IUUCCCCA 881 209 GGGAACAC C CCGCCGCC 506 GGCGGCGG CUGAUGAGGCCGUUAGGCCGAA IUGUUCCC 882 210 GGAACACC C CGCCGCCC 507 GGGCCGCG CUGAUGAGGCCGUUAGGCCGAA IGUGUUCC 883 211 GAACACCC C GCCGCCCU 508 AGGGCGGC CUCAUGAGGCCGUUAGGCCGAA IGGUGUUC 884 214 CACCCCGC C GCCCUCGG 509 CCGAGGGC CUGAUGAGGCCGUUAGGCCGAA ICGGGGUG 885 217 CCCGCCCC C CUCGGAGC 510 GCUCCGAG CUGAUGAGGCCGUUAGGCCGAA ICGGCGGG 886 218 CCGCCGCC C UCGGAGCU 511 AGCUCCGA CUGAUGAGGCCGUUAGGCCGAA IGCGGCGG 887 219 CGCCGCCC U CGGAGCUU 512 AAGCUCCG CUGAUGAGGCCGUUAGGCCGAA IGGCGGCG 888 226 CUCGGAGC U UUUUCUGU 513 ACAGAAAA CUGAUGAGGCCGUUAGGCCGAA ICUCCGAG 889 232 GCUUUUUC U GUGGCGCA 514 UGCGCCAC CUGAUGAGGCCGUUAGGCCGAA IAAAAAGC 890 240 UGUGGCGC A GCUUCUCC 515 GGAGAAGC CUGAUGAGGCCGUUAGGCCGAA ICGCCACA 891 243 GGCGCAGC U UCUCCGCC 516 GGCGGAGA CUGAUGAGGCCGUUAGGCCGAA ICUGCGCC 892 246 GCAGCUUC U CCGCCCGA 517 UCGGGCGG CUGAUGAGGCCGUUAGGCCGAA IAAGCUGC 893 248 AGCUUCUC C GCCCGAGC 518 GCUCGGGC CUGAUGAGGCCGUUAGGCCGAA IAGAAGCU 894 251 UUCUCCGC C CGAGCCGC 519 GCGGCUCG CUGAUGAGGCCGUUAGGCCGAA ICGGAGAA 895 252 UCUCCGCC C GAGCCGCG 520 CGCGGCUC CUGAUGAGGCCGUUAGGCCGAA ICCUGAGA 896 257 GCCCGAGC C GCGCGCGG 521 CCGCGCGC CUGAUGAGGCCGUUAGGCCGAA ICUCGGGC 897 269 CGCGGAGC U GCCGGGGG 522 CCCCCGGC CUGAUGAGGCCGUUAGGCCGAA ICUCCGCG 898 272 GGAGCUGC C GGGGGCUC 523 GAGCCCCC CUGAUGAGGCCGUUAGGCCGAA ICAGCUCC 899 279 CCGGGGGC U CCUUAGCA 524 UGCUAAGG CUGAUGAGGCCGUUAGGCCGAA ICCCCCGG 900 281 GGGGGCUC C UUAGCACC 525 GGUGCUAA CUGAUGAGGCCGUUAGGCCGAA IAGCCCCC 901 282 GGGGCUCC U UAGCACCC 526 GGGUGCUA CUGAUGAGGCCGUUAGGCCGAA IGAGCCCC 902 287 UCCUUAGC A CCCGGGCG 527 CGCCCGGG CUGAUGAGGCCGUUAGGCCGAA ICUAAGGA 903 289 CUUAGCAC C CGGGCGCC 528 GGCGCCCG CUGAUGAGGCCGUUAGGCCGAA IUGCUAAG 904 290 UUAGCACC C GGGCGCCG 529 CGGCGCCC CUGAUGAGGCCGUUAGGCCGAA IGUGCUAA 905 297 CCGGGCGC C GGGGCCCU 530 AGGGCCCC CUGAUGAGGCCGUUAGGCCGAA ICGCCCGG 906 303 GCCGGGGC C CUCGCCCU 531 AGGGCGAG CUGAUGAGGCCGUUAGGCCGAA ICCCCGGC 907 304 CCGGGGCC C UCGCCCUU 532 AAGGGCGA CUGAUGAGGCCGUUAGGCCGAA IGCCCCGG 908 305 CGGGGCCC U CGCCCUUC 533 GAAGGGCG CUGAUGAGGCCGUUAGGCCGAA IGGCCCCG 909 309 GCCCUCGC C CUUCCGCA 534 UGCGGAAG CUGAUGAGGCCGUUAGGCCGAA ICGAGGGC 910 310 CCCUCGCC C UUCCGCAG 535 CUGCGGAA CUGAUGAGGCCGUUAGGCCGAA IGCGAGGG 911 311 CCUCGCCC U UCCGCAGC 536 GCUGCGGA CUGAUGAGGCCGUUAGGCCGAA IGGCGAGG 912 314 CGCCCUUC C GCAGCCUU 537 AAGGCUGC CUGAUGAGGCCGUUAGGCCGAA IAAGGGCG 913 317 CCUUCCGC A GCCUUCAC 538 GUGAAGGC CUGAUGAGGCCGUUAGGCCGAA ICGGAAGG 914 320 UCCGCAGC C UUCACUCC 539 GGAGUGAA CUGAUGAGGCCGUUAGGCCGAA ICUGCGGA 915 321 CCGCAGCC U UCACUCCA 540 UGGAGUGA CUGAUGAGGCCGUUAGGCCGAA IGCUGCGG 916 324 CAGCCUUC A CUCCAGCC 541 GGCUGGAG CUGAUGAGGCCGUUAGGCCGAA IAAGGCUG 917 326 GCCUUCAC U CCAGCCCU 542 AGGGCUGG CUGAUGAGGCCGUUAGGCCGAA IUGAAGGC 918 328 CUUCACUC C AGCCCUCU 543 AGAGGGCU CUGAUGAGGCCGUUAGGCCGAA IAGUGAAG 919 329 UUCACUCC A GCCCUCUG 544 CAGAGGGC CUGAUGAGGCCGUUAGGCCGAA IGAGUGAA 920 332 ACUCCAGC C CUCUGCUC 545 GAGCAGAG CUGAUGAGGCCGUUAGGCCGAA ICUGGAGU 921 333 CUCCAGCC C UCUGCUCC 546 GGAGCAGA CUGAUGAGGCCGUUAGGCCGAA IGCUGGAG 922 334 UCCAGCCC U CUGCUCCC 547 GGGAGCAG CUGAUGAGGCCGUUAGGCCGAA IGGCUGGA 923 336 CAGCCCUC U GCUCCCGC 548 GCGGGAGC CUGAUGAGGCCGUUAGGCCGAA IAGGGCUG 924 339 CCCUCUGC U CCCGCACG 549 CGUGCGGG CUGAUGAGGCCGUUAGGCCGAA ICAGAGGG 925 341 CUCUGCUC C CGCACGCC 550 GGCGUGCG CUGAUGAGGCCGUUAGGCCGAA IAGCAGAG 926 342 UCUGCUCC C GCACGCCA 551 UGGCGUGC CUGAUGAGGCCGUUAGGCCGAA IGAGCAGA 927 345 GCUCCCGC A CGCCAUGA 552 UCAUGGCG CUGAUGAGGCCGUUAGGCCGAA ICGGGAGC 928 349 CCGCACGC C AUGAAGUC 553 GACUUCAU CUGAUGACCCCGUUAGGCCGAA ICGUGCGC 929 350 CGCACGCC A UGAAGUCG 554 CGACUUCA CUGAUGAGGCCGUUAGGCCGAA IGCGUGCG 930 360 GAAGUCGC C GUUCUACC 555 GGUAGAAC CUGAUGAGGCCGUUAGGCCGAA ICGACUUC 931 365 CGCCGUUC U ACCGCUGC 556 GCACCGGU CUGAUGAGGCCGUUAGGCCGAA IAACGGCG 932 368 CGUUCUAC C GCUGCCAG 557 CUGGCAGC CUGAUGAGGCCGUUAGGCCGAA IUAGAACG 933 371 UCUACCUC U GCCAGAAC 558 GUUCUGGC CUGAUGAGGCCGUUAGGCCGAA ICOGUAGA 934 374 ACCGCUGC C AGAACACC 559 GGUGUUCU CUGAUGAGGCCGUUAGGCCGAA ICAGCGGU 935 375 CCGCUGCC A GAACACCA 560 UGGUGUUC CUGAUGAGGCCGUUAGGCCGAA IGCAGCGG 936 380 GCCAGAAC A CCACCUCU 561 AGAGGUGG CUGAUGAGGCCGUUAGGCCGAA IUUCUGGC 937 382 CAGAACAC C ACCUCUGU 562 ACAGAGGU CUGAUGAGGCCGUUAGGCCGAA IUGUUCUG 938 383 AGAACACC A CCUCUGUG 563 CACAGAGG CUGAUGAGGCCGUUAGGCCGAA IGUGUUCU 939 385 AACACCAC C UCUGUGGA 564 UCCACAGA CUGAUGAGGCCGUUAGGCCGAA IUGGUGUU 940 386 ACACCACC U CUGUGGAA 565 UUCCACAG CUGAUGAGGCCGUUAGGCCGAA IGUGGUGU 941 388 ACCACCUC U GUGGAAAA 566 UUUUCCAC CUGAUGAGGCCGUUAGGCCGAA IAGGUGGU 942 401 AAAAAGGC A ACUCGGCG 567 CGCCGAGU CUGAUGAGGCCGUUAGGCCGAA ICCUUUUU 943 404 AAGGCAAC U CGGCGGUG 568 CACCGCCG CUGAUGAGGCCGUUAGGCCGAA IUUGCCUU 944 426 CGGGGUGC U CUUCAGCA 569 UGCUGAAG CUGAUGAGGCCGUUAGGCCGAA ICACCCCG 945 428 GGGUGCUC U UCAGCACC 570 GGUGCUGA CUGAUGAGGCCGUUAGGCCGAA IAGCACCC 946 431 UGCUCUUC A GCACCGGC 571 GCCGGUGC CUGAUGAGGCCGUUAGGCCGAA IAAGAGCA 947 434 UCUUCAGC A CCGGCCUC 572 GAGGCCGG CUGAUGAGGCCGUUAGGCCGAA ICUGAAGA 948 436 UUCAGCAC C GGCCUCCU 573 AGGAGGCC CUGAUGAGGCCGUUAGGCCGAA IUGCUGAA 949 440 GCACCGGC C UCCUGGGC 574 GCCCAGGA CUGAUGAGGCCGUUAGGCCGAA ICCGGUGC 950 441 CACCGGCC U CCUGGGCA 575 UGCCCAGG CUGAUGAGGCCGUUAGGCCGAA IGCCGGUG 951 443 CCGGCCUC C UGGGCAAC 576 GUUGCCCA CUGAUGAGGCCGUUAGGCCGAA IAGGCCGG 952 444 CGGCCUCC U GGGCAACC 577 GGUUGCCC CUGAUGAGGCCGUUAGGCCGAA IGAGGCCG 953 449 UCCUGGGC A ACCUGCUG 578 CAGCAGGU CUGAUGAGGCCGUUAGGCCGAA ICCCAGGA 954 452 UGGGCAAC C UGCUGGCC 579 GGCCAGCA CUGAUGAGGCCGUUAGGCCGAA IUUGCCCA 955 453 GGGCAACC U GCUGGCCC 580 GGGCCAGC CUGAUGAGGCCGUUAGGCCGAA IGUUGCCC 956 456 CAACCUGC U GGCCCUGG 581 CCAGGGCC CUGAUGAGGCCGUUAGGCCGAA ICAGGUUG 957 460 CUGCUGGC C CUGGGGCU 582 AGCCCCAG CUGAUGAGGCCGUUAGGCCGAA ICCAGCAG 958 461 UGCUGGCC C UGGGGCUG 583 CAGCCCCA CUGAUGAGGCCGUUAGGCCGAA IGCCAGCA 959 462 GCUGGCCC U GGGGCUGC 584 GCAGCCCC CUGAUGAGGCCGUUAGGCCGAA IGGCCAGC 960 468 CCUGGGGC U GCUGGCGC 585 GCGCCAGC CUGAUGAGGCCGUUAGGCCGAA ICCCCAGG 961 471 GGGGCUGC U GGCGCGCU 586 AGCGCGCC CUGAUGAGGCCGUUAGGCCGAA ICAGCCCC 962 479 UGGCGCGC U CGGGGCUG 587 CAGCCCCG CUGAUGAGGCCGUUAGGCCGAA ICGCGCCA 963 486 CUCGGGGC U GGGGUGGU 588 ACCACCCC CUGAUGAGGCCGUUAGGCCGAA ICCCCGAG 964 497 GGUGGUGC U CGCGGCGU 589 ACGCCGCG CUGAUGAGGCCGUUAGGCCGAA ICACCACC 965 507 GCGGCGUC C ACUGCGCC 590 GGCGCAGU CUGAUGAGGCCGUUAGGCCGAA IACGCCGC 966 508 CGGCGUCC A CUGCGCCC 591 GGGCGCAG CUGAUGAGGCCGUUAGGCCGAA IGACGCCG 967 510 GCGUCCAC U GCGCCCGC 592 GCGGGCGC CUGAUGAGGCCGUUAGGCCGAA IUGGACGC 968 515 CACUGCGC C CGCUGCCC 593 GGGCAGCG CUGAUGAGGCCGUUAGGCCGAA ICGCAGUG 969 516 ACUGCGCC C GCUGCCCU 594 AGGGCAGC CUGAUGAGGCCGUUAGGCCGAA IGCGCAGU 970 519 GCGCCCGC U GCCCUCGG 595 CCGAGGGC CUGAUGAGGCCGUUAGGCCGAA ICGGGCGC 971 522 CCCGCUGC C CUCGGUCU 596 AGACCGAG CUGAUGAGGCCGUUAGGCCGAA ICAGCGGG 972 523 CCGCUGCC C UCGGUCUU 597 AAGACCGA CUGAUGAGGCCGUUAGGCCGAA IGCAGCGG 973 524 CGCUGCCC U CGGUCUUC 598 GAAGACCG CUGAUGAGGCCGUUAGGCCGAA IGGCAGCG 974 530 CCUCGGUC U UCUACAUG 599 CAUGUAGA CUGAUGAGGCCGUUAGGCCGAA IACCGAGG 975 533 CGGUCUUC U ACAUGCUG 600 CAGCAUGU CUGAUGAGGCCGUUAGGCCGAA IAAGACCG 976 536 UCUUCUAC A UGCUGGUG 601 CACCAGCA CUGAUGAGGCCGUUAGGCCGAA IUAGAAGA 977 540 CUACAUGC U GGUGUGUG 602 CACACACC CUGAUGAGGCCGUUAGGCCGAA ICAUGUAG 978 551 UGUGUGGC C UGACGGUC 603 GACCGUCA CUGAUGAGGCCGUUAGGCCGAA ICCACACA 979 552 GUGUGGCC U GACGGUCA 604 UGACCGUC CUGAUGAGGCCGUUAGGCCGAA IGCCACAC 980 560 UGACGGUC A CCGACUUG 605 CAAGUCGG CUGAUGAGGCCGUUAGGCCGAA IACCGUCA 981 562 ACGGUCAC C GACUUCCU 606 AGCAAGUC CUGAUCAGGCCGUUAGGCCGAA IUGACCGU 982 566 UCACCGAC U UGCUGGGC 607 GCCCAGCA CUGAUGAGGCCGUUAGGCCGAA IUCCGUGA 983 570 CGACUUCC U GCCCAAGU 608 ACUUGCCC CUGAUGAGGCCGUUAGGCCGAA ICAAGUCG 984 575 UGCUGGGC A AGUGCCUC 609 GAGGCACU CUGAUGAGGCCGUUAGGCCGAA ICCCAGCA 985 581 CCAAGUGC C UCCUAAGC 610 GCUUACGA CUGAUGAGGCCGUUAGGCCGAA ICACUUGC 986 582 CAAGUGCC U CCUAAGCC 611 GGCUUAGG CUGAUCACGCCGUUAGGCCGAA IGCACUUG 987 584 AGUGCCUC C UAAGCCCG 612 CGGGCUUA CUGAUGAGGCCGUUAGGCCGAA IAGGCACU 988 585 GUGCCUCC U AAGCCCGG 613 CCGGGCUU CUGAUGAGGCCGUUAGGCCGAA IGAGGCAC 989 590 UCCUAAGC C CGGUGGUG 614 CACCACCG CUGAUGAGGCCGUUAGGCCGAA ICUUAGGA 990 591 CCUAAGCC C CGUGGUGC 615 GCACCACC CUGAUGAGGCCGUUAGGCCGAA IGCUUAGG 991 600 GGUGGUGC U GGCUGCCU 616 AGGCAGCC CUGAUGAGGCCGUUAGGCCGAA ICACCACC 992 604 GUGCUCGC U CCCUACGC 617 GCGUAGGC CUCAUCAGGCCUUUAGGCCGAA ICCAGCAC 993 607 CUGGCUGC C UACGCUCA 618 UGAGCGUA CUGAUGAGGCCGUUAGGCCCAA ICAGCCAG 994 608 UCGCUGCC U ACGCUCAG 619 CUGACCGU CUGAUGAGGCCGUUAGGCCGAA IGCAGCCA 995 613 GCCUACGC U CAGAACCC 620 CGGUUCUG CUGAUGAGGCCGUUAGGCCGAA ICGUAGGC 996 615 CUACGCUC A GAACCGGA 621 UCCGGUUC CUGAUGAGGCCGUUAGGCCGAA IAGCGUAG 997 620 CUCAGAAC C GGAGUCUG 622 CAGACUCC CUGAUGAGGCCGUUAGGCCGAA IUUCUGAG 998 627 CCGGAGUC U GCGGGUGC 623 GCACCCGC CUGAUGAGGCCGUUAGGCCGAA IACUCCGG 999 636 GCCGGUGC U UGCGCCCG 624 CGGGCCCA CUGAUGAGGCCGUUAGGCCGAA ICACCCCC 1000 642 GCUUGCGC C CGCAUUGG 625 CCAAUGCG CUGAUGAGGCCGUUAGGCCGAA ICGCAAGC 1001 643 CUUCCGCC C GCAUUGGA 626 UCCAAUGC CUGAUGAGGCCGUUAGGCCGAA IGCGCAAC 1002 646 GCGCCCGC A UUGGACAA 627 UUGUCCAA CUGAUGAGGCCGUUACGCCGAA ICGGGCGC 1003 653 CAUUGCAC A ACUCGUUG 628 CAACGAGU CUGAUGAGGCCGUUAGGCCGAA IUCCAAUG 1004 656 UGGACAAC U CGUUGUGC 629 GCACAACG CUGAUGAGGCCGUUAGGCCGAA IUUGUCCA 1005 665 CGUUGUGC C AAGCCUUC 630 CAAGCCUU CUGAUGAGGCCGUUAGGCCGAA ICACAACG 1006 666 GUUGUGCC A AGCCUUCG 631 CGAAGGCU CUGAUGAGGCCGUUAGGCCGAA IGCACAAC 1007 670 UGCCAAGC C UUCGCCUU 632 AAGGCGAA CUGAUGAGGCCGUUACGCCCAA ICUUGGCA 1008 671 GCCAAGCC U UCGCCUUC 633 GAACGCGA CUGAUGAGGCCGUUAGGCCGAA IGCUUCGC 1009 676 GCCUUCGC C UUCUUCAU 634 AUGAAGAA CUGAUGAGCCCGUUAGGCCGAA ICGAAGGC 1010 677 CCUUCGCC U UCUUCAUG 635 CAUGAAGA CUGAUGAGGCCGUUAGGCCGAA IGCGAAGG 1011 680 UCGCCUUC U UCAUGUCC 636 GGACAUGA CUGAUGAGGCCGUUAGGCCGAA IAAGGCGA 1012 683 CCUUCUUC A UGUCCUUC 637 GAAGGACA CUGAUGAGGCCGUUAGGCCGAA IAAGAAGG 1013 688 UUCAUGUC C UUCUUUGG 638 CCAAAGAA CUGAUGAGGCCGUUAGGCCGAA IACAUGAA 1014 689 UCAUGUCC U UCUUUGGG 639 CCCAAAGA CUGAUGAGGCCGUUAGGCCGAA IGACAUGA 1015 692 UGUCCUUC U UUGGGCUC 640 GAGCCCAA CUGAUGAGGCCGUUAGGCCGAA IAAGGACA 1016 699 CUUUGGGC U CUCCUCCA 641 UCGAGGAG CUGAUGAGGCCGUUAGGCCGAA ICCCAAAG 1017 701 UUGGGCUC U CCUCGACA 642 UGUCGAGG CUGAUGAGGCCGUUAGGCCGAA IAGCCCAA 1018 703 GGGCUCUC C UCGACACU 643 AGUGUCGA CUGAUGAGGCCGUUAGGCCGAA IAGAGCCC 1019 704 GGCUCUCC U CGACACUG 644 CAGUGUCG CUGAUGAGGCCGUUAGGCCGAA IGAGAGCC 1020 709 UCCUCGAC A CUGCAACU 645 AGUUGCAG CUGAUGAGGCCGUUAGGCCGAA IUCGAGGA 1021 721 CUCGACAC U GCAACUCC 646 GGAGUUGC CUGAUGAGGCCGUUAGGCCGAA IUGUCGAG 1022 714 GACACUGG A ACUCCUGG 647 CCAGGAGU CUGAUGAGGCCGUUAGGCCGAA ICAGUGUC 1023 717 ACUGCAAC U CCUGGCCA 648 UGGCCAGG CUGAUGAGGCCGUUAGGCCGAA IUUGCAGU 1024 719 UGCAACUC C UGGCCAUG 649 CAUGGCCA CUGAUGAGGCCGUUAGGCCGAA IAGUUGCA 1025 720 GCAACUCC U GGCCAUGG 650 CCAUGGCC CUGAUGAGGCCGUUAGGCCGAA IGAGUUGC 1026 724 CUCCUGGC C AUGGCACU 651 AGUGCCAU CUGAUGAGGCCGUUAGGCCGAA ICCAGGAG 1027 725 UCCUGGCC A UGGCACUG 652 CAGUGCCA CUGAUGAGGCCGUUAGGCCGAA IGCCAGGA 1028 730 GCCAUGGC A CUGGAGUG 653 CACUCCAG CUGAUGAGGCCGUUAGGCCGAA ICCAUGGC 1029 732 CAUGGCAC U GGAGUGCU 654 AGCACUCC CUGAUGAGGCCGUUAGGCCGAA IUGCCAUG 1030 740 UCGAGUGC U GGCUCUCC 655 GGAGAGCC CUGAUGAGGCCGUUAGGCCGAA ICACUCCA 1031 744 GUGCUGGC U CUCCCUAG 656 CUAGGGAG CUGAUGAGGCCGUUAGGCCGAA ICCAGCAC 1032 746 GCUGCCUC U CCCUAGGG 657 CCCUAGGG CUGAUGAGGCCGUUAGGCCGAA IAGCCAGC 1033 748 UGGCUCUC C CUAGOCCA 658 UGCCCUAG CUGAUGAGGCCGUUAGGCCGAA IAGAGCCA 1034 749 GGCUCUCC C UAGGGCAC 659 GUCCCCUA CUGAUGAGGCCGUUAGGCCGAA IGAGAGCC 1035 750 GCUCUCCC U AGGGCACC 660 GGUGCCCU CUGAUGAGGCCGUUAGGCCGAA IGGAGAUC 1036 756 CCUAGGGC A CCCUUIJCU 661 AGAAAGGG CUGAUGAGGCCGUUAGGCCGAA ICCCUAGG 1037 758 UAGGGCAC C CUUUCUUC6 62 GAAGAAAG CUGAUGAGGCCGUUAGGCCGAA IUGCCCUA 1038 759 AGGGCACC C UUUCUUCU 663 AGAAGAAA CUGAUGAGGCCGUUAGGCCGAA IGUGCCCU 1039 760 GGGCACCC U UUCUUCUA 664 UAGAAGAA CUGAUGAGGCCGUUAGGCCGAA IGGUGCCC 1040 764 ACCCUUUC U UCUACCGA 665 UCGGUAGA CUGAUGAGGCCGUUAGGCCGAA TAAAGGGU 1041 767 CUUUCUUC U ACCGACGG 666 CCGUCGGU CUGAUGAGGCCGUUAGGCCGAA IAAGAAAG 1042 770 UCUUCUAC C GACGGCAC 667 GUGCCGUC CUGAUGAGCCCGUUAGGCCGAA IUAGAAGA 1043 777 CCGACGGC A CAUCACCC 668 GGGUGAUG CUGAUGAGGCCGUUAGGCCGAA ICCGUCGG 1044 779 GACGGCAC A UCACCCUG 669 CAGGGUGA CUGAUGAGGCCGUUAGGCCGAA IUGCCGUC 1045 782 GGCACAUC A CCCUGCGC 670 GCGCAGGG CUGAUGAGGCCGUUAGGCCGAA IAUGUGCC 1046 784 CACAUCAC C CUGCGCCU 671 AGGCGCAG CUGAUGAGGCCGUUAGGCCGAA TUGAUGUG 1047 785 ACAUCACC C UGCGCCUG 672 CAGGCGCA CUGAUGAGGCCGUUAGGCCGAA IGUGAUGU 1048 786 CAUCACCC U GCGCCUGG 673 CCAGGCGC CUGAUGAGGCCGUUAGGCCGAA IGGUGAUG 1049 791 CCCUGCGC C UGGGCGCA 674 UGCGCCCA CUGAUGAGGCCGUUAGGCCGAA ICGCAGGG 1050 792 CCUGCGCC U GGGCGCAC 675 GUGCGCCC CUGAUGAGGCCGUUAGGCCGAA IGCGCAGG 1051 799 CUGGGCGC A CUGGUGGC 676 GCCACCAG CUGAUGAGGCCGUUAGGCCGAA ICGCCCAG 1052 801 GGGCGCAC U GGUGGCCC 677 GGGCCACC CUGAUGAGGCCGUUAGGCCGAA IUGCGCCC 1053 808 CUGGUGGC C CCGGUGGU 678 ACCACCGG CUGAUGAGGCCGUUAGGCCGAA ICCACCAG 1054 809 UGGUGGCC C CGGUGGUG 679 CACCACCG CUGAUGAGGCCGUUAGGCCGAA IGCCACCA 1055 810 GGUGGCCC C GGUGGUGA 680 UCACCACC CUGAUGAGGCCGUUAGGCCGAA IGGCCACC 1056 823 GUGAGCGC C UUCUCCCU 681 AGGGAGAA CUGAUGAGGCCGUUAGGCCGAA ICOCUCAC 1057 824 UGAGCGCC U UCUCCCUG 682 CAUGGAGA CUGAUGAGGCCGUUAGGCCGAA IGCGCUCA 1058 827 GCGCCUUC U CCCUGGCU 683 AGCCAGGG CUGAUGAGGCCGUUAGGCCGAA IAAGGCGC 1059 829 GCCUUCUC C CUGGCUUU 684 AAAGCCAG CUGAUGAGGCCGUUAGGCCGAA IAGAAGGC 1060 830 CCUUCUCC C UGGCUUUC 685 GAAAGCCA CUGAUGAGGCCGUUAGGCCGAA IGAGAAGG 1061 831 CUUCUCCC U GGCUUUCU 686 AGAAAGCC CUGAUGAGGCCGUUAGGCCGAA IGGAGAAG 1062 835 UCCCUGGC U UUCUGCGC 687 GCGCAGAA CUGAUGAGGCCGUUAGGCCGAA ICCAGGGA 1063 839 UGGCUUUC U GCGCGCUA 688 UAGCGCGC CUGAUGAGGCCGUUAGGCCGAA IAAAGCCA 1064 846 CUGCGCGC U ACCUUUCA 689 UGAAAGGU CUGAUGAGGCCGUUAGGCCGAA ICGCGCAG 1065 849 CGCGCUAC C UUUCAUGG 690 CCAUGAAA CUGAUGAGGCCGUUAGGCCGAA IUAGCGCG 1066 850 GCGCUACC U UUCAUGGG 691 CCCAUGAA CUGAUGAGGCCGUUAGGCCGAA IGUAGCGC 1067 854 UACCUUUC A UGGGCUUC 692 GAAGCCCA CUGAUGAGGCCGUUAGGCCGAA IAAAGGUA 1068 860 UCAUGGGC U UCGGGAAG 693 CUUCCCGA CUGAUGAGGCCGUUAGGCCGAA ICCCAUGA 1069 876 GUUCGUGC A GUACUGCC 694 GGCAGUAC CUGAUGAGGCCGUUAGGCCGAA ICACGAAC 1070 881 UGCAGUAC U GCCCCGGC 695 GCCGGGGC CUGAUGAGGCCGUUAGGCCGAA IUACUGCA 1071 884 AGUACUGC C CCGGCACC 696 GGUGCCGG CUGAUGAGGCCGUUAGGCCGAA ICAGUACU 1072 885 GUACUGCC C CGGCACCU 697 AGGUGCCG CUGAUGAGGCCGUUAGGCCGAA IGCAGUAC 1073 886 UACUGCCC C GGCACCUG 698 CAGGUGCC CUGAUGAGGCCGUUAGGCCGAA IGGCAGUA 1074 890 GCCCCGGC A CCUGGUGC 699 GCACCAGG CUGAUGAGGCCGUUAGGCCGAA ICCGGGGC 1075 892 CCCGGCAC C UGGUGCUU 700 AAGCACCA CUGAUGAGGCCGUUAGGCCGAA IUGCCGGG 1076 893 CCGGCACC U GGUGCLRTU 701 AAAGCACC CUGAUGAGGCCGUUAGGCCGAA IGUGCCGG 1077 899 CCUGGUGC U UUAUCCAG 702 CUGGAUAA CUGAUGAGGCCGUUAGGCCGAA ICACCAGG 1078 905 GCUUUAUC C AGAUGGUC 703 GACCAUCU CUGAUGAGGCCGUUAGGCCGAA IAUAAAGC 1079 906 CUUUAUCC A GAUGGUCC 704 GGACCAUC CUGAUGAGGCCGUUAGGCCGAA IGAUAAAG 1080 914 AGAUGGUC C ACGAGGAG 705 CUCCUCGU CUGAUGAGGCCGUUAGGCCGAA IACCAUCU 1081 915 GAUCGUCC A CCAGGAGG 706 CCUCCUCG CUGAUGAGGCCGUUAGGCCGAA IGACCAUC 1082 926 AGGAGGGC U CGCUGUCG 707 CGACAGCG CUGAUGAGGCCGUUAGGCCGAA ICCCUCCU 1083 930 GGGCUCGC U GUCGGUGC 708 GCACCGAC CUGAUGAGGCCGUUAGGCCGAA ICGAGCCC 1084 939 GUCGGUGC U GGGGUACU 709 AGUACCCC CUGAUGAGGCCGUUAGGCCGAA ICACCGAC 1085 947 UGGGGUAC U CUGUCCUC 710 GAGCACAG CUGAUGAGGCCGUUAGGCCGAA IUACCCCA 1086 949 GOGUACUC U GUGCUCUA 711 UAGAGCAC CUGAUGAGGCCGUUAGGCCGAA IAGUACCC 1087 954 CUCUGUGC U CUACUCCA 712 UGGAGUAG CUGAUGAGGCCGUUAGGCCGAA ICACAGAG 1088 956 CUGUCCUC U ACUCCAGC 713 GCUGGAGU CUGAUGAGGCCGUUAGGCCGAA IAGCACAG 1089 959 UGCUCUAC U CCAGCCUC 714 GAGGCUGG CUGAUGAGGCCGUUAGGCCGAA IUAGAGCA 1090 961 CUCUACUC C AGCCUCAU 715 AUGAGGCU CUGAUGAGGCCGUUAGGCCGAA IAGUAGAG 1091 962 UCUACUCC A GCCUCAUG 716 CAUGAGGC CUGAUGAGGCCGUUAGGCCGAA IGAGUAGA 1092 965 ACUCCAGC C UCAUGGCG 717 CGCCAUGA CUGAUGAGGCCGUUAGGCCGAA ICUGGAGU 1093 966 CUCCAGCC U CAUGGCGC 718 GCGCCAUG CUGAUGAGGCCGUUAGGCCGAA IGCUGGAG 1094 968 CCAGCCUC A UGGCGCUG 719 CAGCGCCA CUGAUGAGGCCGUUAGGCCGAA IAGGCUGG 1095 975 CAUGGCGC U GCUGGUCC 720 GGACCAGC CUGAUGAGGCCGUUAGGCCGAA ICGCCAUG 1096 978 GGCGCUGC U GGUCCUCG 721 CGAGGACC CUGAUGAGGCCGUUAGGCCGAA ICAGCGCC 1097 983 UCCUGGUC C UCGCCACC 722 GGUGGCGA CUGAUGAGGCCGUUAGGCCGAA IACCAGCA 1098 984 GCUGGUCC U CGCCACCG 723 CGGUGGCG CUGAUGAGGCCGUUAGGCCGAA IGACCAGC 1099 988 GUCCUCGC C ACCGUGCU 724 AGCACGGU CUGAUGAGGCCGUUAGGCCGAA ICCAGGAC 1100 989 UCCUCGCC A CCGUGCUG 725 CAGCACGG CUGAUGAGGCCGUUAGGCCGAA IGCGAGGA 1101 991 CUCGCCAC C GUGCUGUG 726 CACAGCAC CUGAUGAGGCCGUUAGGCCGAA IUGGCGAG 1102 996 CACCGUGC U GUGCAACC 727 GGUUGCAC CUGAUGAGGCCGUUAGGCCGAA ICACGGUG 1103 1001 UGCUGUGC A ACCUCGGC 728 GCCGAGGU CUGAUGAGGCCGUUAGGCCGAA ICACAGCA 1104 1004 UGUGCAAC C UCGGCGCC 729 GGCGCCGA CUGAUGAGGCCGUUAGGCCGAA IUUGCACA 1105 1005 GUGCAACC U CGGCGCCA 730 UGGCGCCG CUGAUGAGGCCGUUAGGCCGAA IGUUGCAC 1106 1012 CUCGGCGC C AUGCGCAA 731 UUGCGCAU CUGAUGAGGCCGUUAGGCCGAA ICGCCGAG 1107 1013 UCGGCGCC A UGCGCAAC 732 GUUGCGCA CUGAUGAGGCCGUUAGGCCGAA IGCGCCGA 1108 1019 CCAUGCGC A ACCUCUAU 733 AUAGAGGU CUGAUGAGGCCGUUAGGCCGAA ICGCAUGG 1109 1022 UGCGCAAC C UCUAUGCG 734 CGCAUAGA CUGAUGAGGCCGUUAGGCCGAA IUUGCGCA 1110 1023 GCGCAACC U CUAUGCGA 735 UCGCAUAG CUGAUGAGGCCGUUAGGCCGAA IGUUGCGC 1111 1025 GCAACCUC U AUGCGAUG 736 CAUCGCAU CUGAUGAGGCCGUUAGGCCGAA IAGGUUGC 1112 1035 UGCGAUGC A CCGGCGGC 737 GCCGCCGG CUGAUGAGGCCGUUAGGCCGAA ICAUCGCA 1113 1037 CGAUGCAC C GGCGGCUG 738 CAGCCGCC CUGAUGAGGCCGUUAGGCCGAA IUGCAUCG 1114 1044 CCGGCGGC U GCAGCGGC 739 GCCGCUGC CUGAUGAGGCCGUUAGGCCGAA ICCGCCGG 1115 1047 GCGGCUGC A GCGGCACC 740 GGUGCCGC CUGAUGAGGCCGUUAGGCCGAA ICAGCCGC 1116 1053 GCAGCGGC A CCCGCGCU 741 AGCGCGGG CUGAUGAGGCCGUUAGGCCGAA ICCGCUGC 1117 1055 AGCGGCAC C CGCGCUCC 742 GGAGCGCG CUGAUGAGGCCGUUAGGCCGAA IUGCCGCU 1118 1056 GCGGCACC C GCGCUCCU 743 AGGAGCGC CUGAUGAGGCCGUUAGGCCGAA IGUGCCGC 1119 1061 ACCCGCGC U CCUGCACC 744 GGUGCAGG CUGAUGAGGCCGUUAGGCCGAA ICGCGGGU 1120 1063 CCGCGCUC C UCCACCAG 745 CUGGUCCA CUGAUGAGGCCGUUAGGCCGAA IAGCGCGG 1121 1064 CGCGCUCC U GCACCAGG 746 CCUGGUGC CUGAUGAGGCCGUUAGGCCGAA IGAGCGCG 1122 1067 GCUCCUGC A CCAGGGAC 747 GUCCCUGG CUGAUGAGGCCGUUAGGCCGAA ICAGGAGC 1123 1069 UCCUCCAC C AGGGACUG 748 CAGUCCCU CUGAUGAGGCCGUUAGGCCGAA IUCCAGGA 1124 1070 CCUGCACC A GGGACUGU 749 ACAGUCCC CUGAUGAGGCCGUUAGGCCGAA IGUGCAGG 1125 1076 CCAGGGAC U GUGCCGAG 750 CUCGGCAC CUGAUGAGGCCGUUAGGCCGAA IUCCCUGG 1126 1081 GACUGUGC C GAGCCGCG 751 CGCGGCUC CUGAUGAGGCCGUUAGGCCGAA ICACAGUC 1127 1086 UGCCGAGC C GCGCGCGG 752 CCGCGCGC CUGAUGAGGCCGUUAGGCCGAA ICUCGGCA 1128 1111 GAAGCGUC C CCUCAGCC 753 GGCUGAGG CUGAUGAGGCCGUUAGGCCGAA IACGCUUC 1129 1112 AAGCGUCC C CUCAGCCC 754 GGGCUGAG CUGAUGAGGCCGUUAGGCCGAA IGACGCUU 1130 1113 AGCGUCCC C UCAGCCCC 755 GGGGCUGA CUGAUGAGGCCGUUAGGCCGAA IGGACGCU 1131 1114 GCGUCCCC U CAGCCCCU 756 AGGGGCUG CUGAUGAGGCCGUUAGGCCGAA IGGGACGC 1132 1116 GUCCCCUC A GCCCCUGG 757 CCAGGGGC CUGAUGAGGCCGUUAGGCCGAA IAGGGGAC 1133 1119 CCCUCAGC C CCUCGAGG 758 CCUCCAGG CUGAUGACGCCGUUAGGCCGAA ICUGAGGG 1134 1120 CCUCAGCC C CUGGAGGA 759 UCCUCCAG CUGAUGAGGCCGUUAGGCCGAA IGCUGAGG 1135 1121 CUCAGCCC C UGGAGGAG 760 CUCCUCCA CUGAUGAGGCCGUUAGGCCGAA IGGCUGAG 1136 1122 UCAGCCCC U GGAGGAGC 761 OCUCCUCO CUGAUGAGGCCGUUAGGCCGAA IGGGCUGA 1137 1131 GGAGGAGC U GGAUCACC 762 GGUGAUCC CUGAUGAGGCCGUUAGGCCGAA ICUCCUCC 1138 1137 GCUGGAUC A CCUCCUGC 763 GCAGGAGG CUGAUGAGGCCGUUAGGCCGAA IAUCCAGC 1139 1139 UGGAUCAC C UCCUGCUG 764 CAGCAGGA CUGAUGAGGCCGUUAGGCCGAA IUGAUCCA 1140 1140 GGAUCACC U CCUGCUGC 765 GCAGCAGG CUGAUGAGGCCGUUAGGCCGAA TGUGAUCC 1141 1142 AUCACCUC C UGCUGCUG 766 CAGCAGCA CUGAUGAGGCCGUUAGGCCGAA IAGGUGAU 1142 1143 UCACCUCC U GCUGCUGG 767 CCAGCAGC CUGAUGAGGCCGUUAGGCCGAA IGAGGUGA 1143 1146 CCUCCUGC U GCUGGCGC 768 GCGCCAGC CUGAUGAGGCCGUUAGGCCGAA ICAGGAGG 1144 1149 CCUGCUGC U GGCGCUGA 769 UCAGCGCC CUGAUGAGGCCGUUAGGCCGAA ICAGCAGG 1145 1155 GCUGGCGC U GAUGACCG 770 CGGUCAUC CUGAUGAGGCCGUUAGGCCGAA ICGCCAGC 1146 1162 CUGAUGAC C GUGCUCUU 771 AAGAGCAC CUGAUGAGGCCGUUAGGCCGAA IUCAUCAG 1147 1167 GACCGUGC U CUUCACUA 772 UAGUGAAG CUGAUGAGGCCGUUAGGCCGAA ICACGGUC 1148 1169 CCGUGCUC U UCACUAUG 773 CAUAGUGA CUGAUGAGGCCGUUAGGCCGAA IAGCACGG 1149 1172 UGCUCUUC A CUAUGUGU 774 ACACAUAG CUGAUGAGGCCGUUAGGCCGAA IAAGAGCA 1150 1174 CUCUUCAC U AUGUGUUC 775 GAACACAU CUGAUGAGGCCGUUAGGCCGAA IUGAAGAG 1151 1183 AUGUGUUC U CUGCCCGU 776 ACGGGCAG CUGAUGAGGCCGUUAGGCCGAA IAACACAU 1152 1185 GUGUUCUC U GCCCGUAA 777 UUACGGGC CUGAUGAGGCCGUUAGGCCGAA IAGAACAC 1153 1188 UUCUCUGC C CGUAAUUU 778 AAAUUACG CUGAUGAGGCCGUUAGGCCGAA ICAGAGAA 1154 1189 UCUCUGCC C GUAAUUUA 779 UAAAUUAC CUGAUGAGGCCGUUAGGCCGAA IGCAGAGA 1155 1204 UAUCGCGC U UACUAUGG 780 CCAUAGUA CUGAUGAGGCCGUUAGGCCGAA ICGCGAUA 1156 1208 GCGCUUAC U AUGGAGCA 781 UGCUCCAU CUGAUGAGGCCGUUAGGCCGAA IUAAGCGC 1157 1216 UAUGGAGC A UUUAAGGA 782 UCCUUAAA CUGAUGAGGCCGUUAGGCCGAA ICUCCAUA 1158 1229 AGGAUGUC A AGGAGAAA 783 UUUCUCCU CUGAUGAGGCCGUUAGGCCGAA IACAUCCU 1159 1241 AGAAAAAC A GGACCUCU 784 AGAGGUCC CUGAUGAGGCCGUUAGGCCGAA IUUUUUCU 1160 1246 AACAGGAC C UCUGAAGA 785 UCUUCAGA CUGAUGAGGCCGUUAGGCCGAA IUCCUGUU 1161 1247 ACAGGACC U CUGAAGAA 786 UUCUUCAG CUGAUGAGGCCGUUAGGCCGAA IGUCCUGU 1162 1249 AGGACCUC U GAAGAAGC 787 GCUUCUUC CUGAUGAGGCCGUUAGGCCGAA IAGGUCCU 1163 1258 GAAGAAGC A GAAGACCU 788 AGGUCUUC CUGAUGAGGCCGUUAGGCCGAA ICUUCUUC 1164 1265 CAGAAGAC C UCCGAGCC 789 GGCUCGGA CUGAUGAGGCCGUUAGGCCGAA IUCUUCUG 1165 1266 AGAAGACC U CCGAGCCU 790 AGGCUCGG CUGAUGAGGCCGUUAGGCCGAA IGUCUUCU 1166 1268 AAGACCUC C GAGCCUUG 791 CAAGGCUC CUGAUGAGGCCGUUAGGCCGAA IAGGUCUU 1167 1273 CUCCGAGC C UUGCGAUU 792 AAUCGCAA CUGAUGAGGCCGUUAGGCCGAA ICUCGGAG 1168 1274 UCCGAGCC U UUGCGAUU 793 AAAUCGCA CUGAUGAGGCCGUUAGGCCGAA IGCUCGGA 1169 1284 GCGAUUUC U AUCUGUGA 794 UCACAGAU CUGAUGAGGCCGUUAGGCCGAA IAAAUCGC 1170 1288 UUUCUAUC U GUGAUUUC 795 GAAAUCAC CUGAUGAGGCCGUUAGGCCGAA IAUAGAAA 1171 1297 GUGAUUUC A AUUGUGGA 796 UCCACAAU CUGAUGAGGCCGUUAGGCCGAA IAAAUCAC 1172 1307 UUGUGGAC C CUUGGAUU 797 AAUCCAAG CUGAUGAGGCCGUUAGGCCGAA IUCCACAA 1173 1308 UGUGGACC C UUGGAUUU 798 AAAUCCAA CUGAUGAGGCCGUUAGGCCGAA IGUCCACA 1174 1309 GUGGACCC U UGGAUUUU 799 AAAAUCCA CUGAUGAGGCCGUUAGGCCGAA IGGUCCAC 1175 1322 UUUUUAUC A UUUUCAGA 800 UCUGAAAA CUGAUGAGGCCGUUAGGCCGAA IAUAAAAA 1176 1328 UCAUUUUC A GAUCUCCA 801 UGGAGAUC CUGAUGAGGCCGUUAGGCCGAA IAAAAUGA 1177 1333 UUCAGAUC U CCAGUAUU 802 AAUACUGG CUGAUGAGGCCGUUAGGCCGAA IAUCUGAA 1178 1335 CAGAUCUC C AGUAUUUC 803 GAAAUACU CUGAUGAGGCCGUUAGGCCGAA IAGAUCUG 1179 1336 AGAUCUCC A GUAUUUCG 804 CGAAAUAC CUGAUGAGGCCGUUAGGCCGAA IGAGAUCU 1180 1356 AUUUUUUC A CAAGAUUU 805 AAAUCUUG CUGAUGAGGCCGUUAGGCCGAA IAAAAAAU 1181 1358 UUUUUCAC A AGAUUUUC 806 GAAAAUCU CUGAUGAGGCCGUUAGGCCGAA IUGAAAAA 1182 1367 AGAUUUUC A UUAGACCU 807 AGGUCUAA CUGAUGAGGCCGUUAGGCCGAA IAAAAUCU 1183 1374 CAUUACAC C UCUUAGGU 808 ACCUAAGA CUGAUGAGGCCGUUAGCCCGAA IUCUAAUG 1184 1375 AUUAGACC U CUUAGGUA 809 UACCUAAG CUGAUGAGGCCGUUAGGCCGAA IGUCUAAU 1185 1377 UAGACCUC U UAGGUACA 810 UGUACCUA CUGAUGAGGCCGUUAGGCCGAA IAGGUCUA 1186 1385 UUAGGUAC A GGAGCCGG 811 CCGGCUCC CUGAUGAGGCCGUUAGGCCGAA IUACCUAA 1187 1391 ACAGGAGC C GGUGCAGC 812 GCUGCACC CUGAUGAGGCCGUUAGGCCGAA ICUCCUGU 1188 1397 GCCGGUGC A GCAAUUCC 813 GGAAUUGC CUGAUGAGGCCGUUAGGCCGAA ICACCGGC 1189 1400 GGUGCAGC A AUUCCACU 814 AGUGCAAU CUGAUGAGGCCGUUAGGCCGAA ICUGCACC 1190 1405 AGCAAUUC C ACUAACAU 815 AUGUUAGU CUGAUGAGGCCGUUAGGCCGAA IAAUUGCU 1191 1406 GCAAUUCC A CUAACAUG 816 CAUGUUAG CUGAUGAGGCCGUUAGCCCGAA IGAAUUGC 1192 1408 AAUUCCAC U AACAUGGA 817 UCCAUGUU CUGAUGAGGCCGUUAGGCCGAA IUGGAAUU 1193 1412 CCACUAAC A UGGAAUCC 818 GGAUUCCA CUGAUGAGGCCGUUAGGCCGAA IUUAGUGG 1194 1420 AUCGAAUC C AGUCUGUG 819 CACAGACU CUGAUGAGGCCGUUAGGCCGAA IAUUCCAU 1195 1421 UGGAAUCC A GUCUGUGA 820 UCACAGAC CUGAUGAGGCCGUUAGGCCGAA IGAUUCCA 1196 1425 AUCCAGUC U GUGACAGU 821 ACUGUCAC CUGAUGAGGCCGUUAGGCCGAA IACUGGAU 1197 1431 UCUGUGAC A GUGUUUUU 822 AAAAACAC CUGAUGAGGCCGUUAGGCCGAA IUCACAGA 1198 1441 UGUUUUUC A CUCUGUGG 823 CCACAGAG CUGAUGAGGCCGUUAGGCCGAA IAAAAACA 1199 1443 UUUUUCAC U CUGUGGUA 824 UACCACAG CUGAUGAGGCCGUUAGGCCGAA IUGAAAAA 1200 1445 UUUCACUC U GUGGUAAG 825 CUUACCAC CUGAUGAGGCCGUUAGGCCGAA IAGUGAAA 1201 1455 UGGUAAGC U GAGGAAUA 826 UAUUCCUC CUGAUGAGGCCGUUAGGCCGAA ICUUACCA 1202 1468 AAUAUGUC A CAUUUUCA 827 UGAAAAUG CUGAUGAGGCCGUUAGGCCGAA IACAUAUU 1203 1470 UAUGUCAC A UUUUCAGU 828 ACUGAAAA CUGAUGAGGCCGUUAGGCCGAA IUGACAUA 1204 1476 ACAUUUUC A GUCAAAGA 829 UCUUUGAC CUGAUGAGGCCGUUAGGCCGAA IAAAAUGU 1205 1480 UUUCAGUC A AAGAACCA 830 UGGUUCUU CUGAUGAGGCCGUUAGGCCGAA IACUGAAA 1206

[0232] TABLE V Human PTGDR Zinzyme and Substrate Sequence Seq Seq Pos Substrate ID Zinzyme ID 9 GAAUUCUG G CUAUUUUC 1207 GAAAAUAG GCCGAAAGGCGAGUGAGGUCU CAGAAUUC 1438 23 UUCCUCCU G CCGUUCCG 1208 CGGAACGG GCCGAAAGGCGAGUGAGGUCU AGGAGGAA 1439 26 CUCCUGCC G UUCCGACU 1209 AGUCGGAA GCCGAAAGGCGAGUGAGGUCU GGCAGGAG 1440 37 CCGACUCG G CACCAGAG 1210 CUCUGGUG GCCGAAAGGCGAGUGAGGUCU CGAGUCGG 1441 45 GCACCAGA G UCUGUCUC 1211 GAGACAGA GCCGAAAGGCGAGUGAGGUCU UCUGGUGC 1442 49 CAGAGUCU G UCUCUACU 1212 AGUAGAGA GCCGAAAGGCGAGUGAGGUCU AGACUCUG 1443 64 CUGAGAAC G CAGCGCGU 1213 ACGCGCUG GCCGAAAGGCGAGUGAGGUCU GUUCUCAG 1444 67 AGAACGCA G CGCGUCAG 1214 CUGACGCG GCCGAAAGGCGAGUGAGGUCU UGCGUUCU 1445 69 AACGCAGC G CGUCAGGG 1215 CCCUGACG GCCGAAAGGCGAGUGAGGUCU GCUGCGUU 1446 71 CGCAGCGC G UCAGGGCC 1216 GGCCCUGA GCCGAAAGGCGAGUGAGGUCU GCGCUGCG 1447 77 GCGUCAGG G CCGAGCUC 1217 GAGCUCGG GCCGAAAGGCGAGUGAGGUCU CCUGACGC 1448 82 AGGGCCGA G CUCUUCAC 1218 GUGAAGAG GCCGAAAGGCGAGUGAGGUCU UCGGCCCU 1449 93 CUUCACUG G CCUGCUCC 1219 GGAGCAGG GCCGAAAGGCGAGUGAGGUCU CAGUGAAG 1450 97 ACUGGCCU G CUCCUCUC 1220 GCGCGGAG GCCGAAAGGCGAGUGAGGUCU AGGCCAGU 1451 102 CCUGCUCC G CGCUCUUC 1221 GAAGAGCG GCCGAAAGGCGAGUGAGGUCU GGAGCAGG 1452 104 UGCUCCGC G CUCUUCAA 1222 UUGAAGAG GCCGAAAGGCGAGUGAGGUCU GCGGAGCA 1453 114 UCUUCAAU G CCAGCGCC 1223 GGCGCUGG GCCUAAAGGCGAGUGAGGUCU AUUGAAUA 1454 118 CAAUGCCA G CGCCAGGC 1224 GCCUGGCG GCCGAAAGGCGAUUGAGGUCU UGGCAUUU 1455 120 AUGCCAUC G CCAGUCGC 1225 GCGCCUGG GCCGAAAGGCGAGUGAGGUCU GCUGGCAU 1456 125 AUCUCCAG G CGCUCACC 1226 GGUGAGCG GCCGAAAGGCGAGUGAGGUCU CUGGCGCU 1457 127 CGCCAGGC G CUCACCCU 1227 AUGGUGAG GCCGAAAGGCGAGUUAGGUCU GCCUGGCG 1458 136 CUCACCCU G CAGAGCGU 1228 ACUCUCUG GCCGAAAGGCGAGUGAGGUCU AGUGUGAG 1459 141 CCUUCAUA G CGUCCCGC 1229 UCUUGACU GCCGAAAGGCGAGUGAGGUCU UCUGCAGG 1460 143 UUCAGAGC G UCCCUCCU 1230 AGUCUGGA GCCGAAAGGCGAGUGAGGUCU UCUCUUCA 1461 148 AUCUUCCC G CCUCUCAA 1231 UUGAGAGG GCCGAAAUGCGAGUGAGGUCU GGGACGCU 1462 163 AAAGAGGG G UGUGACCC 1232 GUGUCACA GCCGAAAGGCGAGUGAGGUCU CCCUCUUU 1463 165 AUAUGUGU G UGACCCGC 1233 UCUGGUCA GCCUAAAGUCGAGUGAGGUCU ACCCCUCU 1464 172 UGUGACCC G CGAGUUUA 1234 UAAACUCG GCCGAAAGGCGAGUGAGGUCU GUGUCACA 1465 176 ACCCGCGA G UUUAGAUA 1235 UAUCUAAA GCCGAAAGGCGAGUGAGGUCU UCUCUUGU 1466 189 UAUAUUAG G UUCCUGCC 1236 GGCAGGAA GCCGAAAUGCGAGUGAGGUCU CUCCUAUC 1467 195 AGGUUCCU G CCUUUUUU 1237 CCCCACUU GCCGAAAGGCGAGUGAGGUCU AGGAACCU 1468 198 UUCCUGCC G UGGGGAAC 1238 GUUCCCCA GCCGAAAGUCUAGUGAGGUCU GGCAGGAA 1469 212 AACACCCC G CCUCCCUC 1239 GAGGUCUG GCCGAAAGGCGAGUGAGGUCU GGGGUGUU 1470 215 ACCCCGCC G CCCUCGGA 1240 UCCUAGUG GCCGAAAUUCUAGUGAGGUCU UUCUUUUU 1471 224 CCCUCGGA G CUUUUUCU 1241 AGAAAAAU UCCGAAAGGCGAGUGAUGUCU UCCGAGGG 1472 233 CUUUUUCU G UGUCUCAG 1242 CUGCGCCA UCCGAAAGGCGAGUGAGGUCU AGAAAAAG 1473 236 UUUCUGUG G CUCAUCUU 1243 AAUCUUCU UCCUAAAGGCUAUUGAUUUCU CACAGAAA 1474 238 UCUGUGUC G CAGCUUCU 1244 AGAAGCUG UCCUAAAUUCUAUUUAGGUCU UCCACAGA 1475 241 GUGUCUCA G CUUCUCCG 1245 CUUAUAAU GCCGAAAGGCUAUUUAUGUCU UGCGCCAC 1476 249 GCUUCUCC G CCCUAUCC 1246 GGCUCUUU UCCUAAAUUCGAGUUAGUUCU UGAUAAUC 1477 255 CCUCCCGA G CCGCUCUC 1247 UCGCGCUG UCCUAAAUUCUAUUUAUGUCU UCUUUCUU 1478 258 CCCUAUCC G CUCGCGGA 1248 UCCUCGCU GCCGAAAGUCGAUUUAUUUCU GUCUCGGG 1479 260 CGAGCCGC G CUCUUAUC 1249 GCUCCGCU UCCUAAAUUCUAGUGAUUUCU UCGUCUCU 1480 262 AUCCUCUC G CUUAGCUG 1250 CAGCUCCU GCCUAAAGGCGAUUUAUGUCU UCUCGGCU 1481 267 CGCGCGGA G CUUCCUUU 1251 CCCGGCAU UCCUAAAUUCUAGUGAUUUCU UCCUCUCU 1482 270 UCUUAUCU G CCGGGGGC 1252 UCCCCCUG GCCUAAAUUCUAUUUAUUUCU AUCUCCUC 1483 277 UGCCGGUU G CUCCUUAU 1253 CUAAUUAU UCCUAAAUUCGAGUGAUUUCU CCCCGUCA 1484 285 GCUCCUUA G CACCCGGG 1254 CCCGGGUG GCCGAAAGGCGAGUGAGGUCU UAAGGAGC 1485 293 GCACCCGG G CGCCGGGG 1255 CCCCGGCG GCCGAAAGGCGAGUGAGGUCU CCGGGUGC 1486 295 ACCCGGGC G CCGGGGCC 1256 GGCCCCGG GCCGAAAGGCGAGUGAGGUCU GCCCGGGU 1487 301 GCGCCGGG G CCCUCGCC 1257 GGCGAGGG GCCGAAAGGCGAGUCAGGUCU CCCGGCGC 1488 307 GGGCCCUC G CCCUUCCG 1258 CGGAAGGG GCCGAAAGGCGAGUGAGGUCU GAGGGCCC 1489 315 GCCCUUCC G CAGCCUUC 1259 GAAGGCUG GCCGAAAGGCGAGUGAGGUCU GGAAGGGC 1490 318 CUUCCGCA G CCUUCACU 1260 AGUGAAGG CCCGAAAGGCGAGUGAGGUCU UGCCGAAG 1491 330 UCACUCCA G CCCUCUGC 1261 GCAGAGGG GCCGAAAGGCGAGUGAGGUCU UGGACUGA 1492 337 AGCCCUCU G CUCCCGCA 1262 UGCGGGAG GCCGAPAGGCGAGUGAGGUCU AGAGGGCU 1493 343 CUGCUCCC G CACGCCAU 1263 AUGGCGUG GCCGAAAGGCGAGUGAGGUCU GGGAGCAC 1494 347 UCCCGCAC G CCAUGAAG 1264 CUUCAUGG GCCGAAAGGCGAGUGAGGUCU GUGCGGGA 1495 355 GCCAUGAA G UCGCCGUU 1265 AACGGCCA CCCGAAAGGCGAGUGAGGUCU UUCAUGGC 1496 358 AUGAAGUC G CCGUUCUA 1266 UAGAACGG GCCGAAAGCCGAGUGAGGUCU GACUUCAU 1497 361 AAGUCGCC G UUCUACCG 1267 CGGUACAA CCCGAAAGGCGAGUGAGCUCU GGCGACUU 1498 369 GUUCUACC G CUCCOAGA 1268 UCUGGCAG GCCGAAACGCGAGUGAGGUCU CGUAGAAC 1499 372 CUACCGCU G CCAGAACA 1269 UGUUCUCC GCCGAAAGGCGACUCACCUCU AGCCGUAC 1500 389 CCACCUCU G UGGAAAAA 1270 UUUUUCCA GCCCAAAGGCGAGUGAGGUCU AGAGGUOG 1501 399 GGAAAAAG G CAACUCGG 1271 CCCAGUUG GCCGAAAGGCGAGUGAGGUCU CUUUUUCC 1502 407 GCAACUCG G CUCUGAUC 1272 CAUCACCC GCCCAAAGGCGAGUGACCUCU CGAGUUGC 1503 410 ACUCGGCC G UGAUGGUC 1273 GCCCAUCA GCCGAAACCCGAGUGAGGUCU CGCCGAGU 1504 417 GGUGAUGG G CGCCCUGC 1274 CCACCCCG GCCGAAAGGCCAGUCACCUCU CCAUCACC 1505 422 UGGGCCGC G UGCUCUUC 1275 GAAGAGCA GCCCAAAGGCGAGUGAGCUCU CCCGCCCA 1506 424 CGCGGGGU G CUCUUCAG 1276 CUGAAGAG CCCGAAAGGCGAGUGAGGUCU ACCCCCCC 1507 432 GCUCUUCA G CACCGGCC 1277 GGCCGGUG CCCGAAAGGCGAGUGAGGUCU UCAAGAGC 1508 438 CAGCACCG G CCUCCUCC 1278 CCAGCAGG GCCGAAAGGCGAGUGAGGUCU CCGUGCUG 1509 447 CCUCCUCG G CAACCUGC 1279 GCAGGUUG GCCCAAAGGCGAGUGAGCUCU CCAGGAGG 1510 454 GCCAACCU G CUCGCCCU 1280 AGGGCCAG CCCGAAAGGCGAGUCAGGUCU AGGUUCCC 1511 458 ACCUCCUC G CCCUGGGG 1281 CCCCAGGC GCCCAAAGGCGAGUGACGUCU CAGCAGGU 1512 466 GCCCUGGG G CUGCUCGC 1282 GCCAGCAG GCCGAAAGGCCAGUGAGGUCU CCCAGCCC 1513 469 CUGGGGCU G CUGGCCCG 1283 CGCGCCAG GCCGAAAGGCGAGUCAGGUCU AGCCCCAC 1514 473 GGCUGCUG G CGCCCUCG 1284 CGAGCGCG GCCGAAAGGCGAGUGAGGUCU CACCAGCC 1515 475 CUGCUGGC G CCCUCCGG 1285 CCCCACCC GCCGAAACCCCACUCAGGUCU GCCACCAC 1516 477 CCUCGCGC G CUCGGCCC 1286 CCCCCCAC CCCCAAACGCGAGUCACCUCU CCGCCACC 1517 484 CCCUCCCC G CUCCGGUG 1287 CACCCCAG GCCCAAACCCCACUCAGCUCU CCCCACCC 1518 490 CCCCUGCG G UGGUCCUC 1288 CACCACCA CCCCAAACCCCACUCACCUCU CCCACCCC 1519 493 CUGCCCUC G UCCUCCCG 1289 CCCGACCA GCCCAAACCCCAGUCACCUCU CACCCCAG 1520 495 CCCCUCGU G CUCCCCCC 1290 CCCCCCAC CCCCAAAGCCGACUCACCUCU ACCACCCC 1521 499 UCCUCCUC G CCCCCUCC 1291 CCACGCCG CCCGAAACCCCACUGAGCUCU CACCACCA 1522 502 UCCUCGCG G CCUCCACU 1292 AGUCCACC CCCGAAAGGCGACUCACCUCU CGCCACCA 1523 504 CUCCCCCC G UCCACUCC 1293 GCACUCCA CCCCAAACGCGAGUCACGUCU CCCCCGAC 1524 511 CCUCCACU G CCCCCCCU 1294 ACCCCGCG GCCGAAACCCCACUCAGGUCU AGUCCACC 1525 513 UCCACUGC G CCCCCUCC 1295 CCACCCCC CCCCAAACCCCACUCACCUCU CCAGUCCA 1526 517 CUCCGCCC G CUGCCCUC 1296 CACGCCAC GCCCAAACCCGAGUCAGGUCU GGGCCCAC 1527 520 CGCCCGCU G CCCUCCGU 1297 ACCGACCG GCCGAAAGGCCAGUCACCUCU AGCCCCCC 1528 527 UGCCCUCG G UCUUCUAC 1298 GUACAACA GCCGAAACGCGAGUGAGGUCU CCACGCCA 1529 538 UUCUACAU G CUCCUCUC 1299 CACACCAG CCCGAAAGGCGAGUCAGGUCU AUCUACAA 1530 542 ACAUCCUC G UGUGUGCC 1300 GCCACACA GCCGAAACCCGACUGAGGUCU CACCAUGU 1531 544 AUCCUCCU G UGUCCCCU 1301 AGCCCACA GCCGAAACCCGAGUCACGUCU ACCACCAU 1532 546 GCUCCUGU G UGCCCUGA 1302 UCACGCCA CCCCAAACCCCAGUGACCUCU ACACCACC 1533 549 CCUGUCUG G CCUCACCC 1303 CCCUCACG CCCCAAACCCGACUCAGGUCU CACACACC 1534 557 GCCUGACC G UCACCGAC 1304 CUCGGUGA CCCCAAACCCGAGUGACGUCU CCUCACCC 1535 568 ACCCACUU G CUGGGCAA 1305 UUGCCCAG GCCGAAAGGCGAGUGAGGUCU AAGUCGGU 1536 573 CUUGCUGG G CAAGUGCC 1306 GGCACUUG GCCGAAAGGCGAGUGAGGUCU CCAGCAAG 1537 577 CUGGGCAA G UGCCUCCU 1307 AGGAGGCA GCCGAAAGGCGAGUGAGGUCU UUGCCCAG 1538 579 GGGCAAGU G CCUCCUAA 1308 UUAGGAGG GCCGAAAGGCGAGUGAGGUCU ACUUGCCC 1539 588 CCUCCUAA G CCCGGUGG 1309 CCACCGGG GCCGAAAGGCGAGUGAGGUCU UUAGGACG 1540 593 UAAGCCCG G UGGUGCUG 1310 CAGCACCA GCCGAAAGGCGAGUGAGGUCU CGGGCUUA 1541 596 GCCCGGUG G UGCUGGCU 1311 AGCCAGCA GCCGAAAGGCGAGUGAGGUCU CACCGGGC 1542 598 CCGGUGGU G CUGGCUGC 1312 GCAGCCAG GCCGAAAGGCGAGUGAGGUCU ACCACCGG 1543 602 UGGUGCUG G CUOCCUAC 1313 GUAGUCAG GCCGAAAGGCGAGUGAGGUCU CAGCACCA 1544 605 UGCUGGCU G CCUACGCU 1314 AGCGUAGG GCCGAAAGGCGAGUGAGGUCU AGCCAGCA 1545 611 CUGCCUAC G CUCAGAAC 1315 GUUCUGAG GCCGAAAGGCGAGUGAGGUCU GUAGGCAG 1546 624 GAACCGGA G UCUGCGGG 1316 CCCGCAGA GCCGAAAGGCGAGUGAGGUCU UCCGCUUC 1547 628 CGGAGUCU G CGGGUGCU 1317 AGCACCCG GCCGAAAGGCGAGUGAGCUCU AGACUCCG 1548 632 GUCUGCGG G UGCUUGCG 1318 CGCAAGCA GCCGAAAGGCGAGUGAGGUCU CCGCAGAC 1549 634 CUGCGGGU G CUUGCGCC 1319 GGCGCAAG GCCGAAAGGCGAGUGAGGUCU ACCCGCAG 1550 638 GGGUGCUU G CGCCCGCA 1320 UGCGGGCG GCCGAAAGGCGAGUGAGGUCU AAGCACCC 1551 640 GUGCUUGC G CCCGCAUU 1321 AAUGCGGG GCCGAAAGGCGAGUGAGGUCU GCAAGCAC 1552 644 UUGCGCCC G CAUUGGAC 1322 GUCCAAUG GCCGAAAGGCGAGUGAGGUCU GGGCGCAA 1553 658 GACAACUC G UUGUGCCA 1323 UGGCACAA GCCGAAAGGCGAGUGAGGUCU GAGUGGUC 1554 661 AACUCGUU G UGCCAAGC 1324 GCUUGGCA GCCGAAAGGCGAGUGAGGUCU AACGAGUU 1555 663 CUCGUUGU G CCAAGCCU 1325 AGGCUUGG GCCGAAAGGCGAGUGAGGUCU ACAACGAG 1556 668 UGUGCCAA G CCUUCGCC 1326 GGCGAAGG GCCGAAAGGCGAGUGAGGUCU UUGGCACA 1557 674 AAGCCUUC G CCUUCUUC 1327 GAAGAAGG GCCGAAAGGCGAGUGAGGUCU GAAGGCUU 1558 685 UUCUUCAU G UCCUUCUU 1328 AAGAAGGA GCCGAAAGGCGAGUGAGGUCU AUGAAGAA 1559 697 UUCUUUGG G CUCUCCUC 1329 GAGGAGAG GCCGAAAGGCGAGUGAGGUCU CCAAAGAA 1560 712 UCGACACU G CAACUCCU 1330 AGGAGUUG GCCGAAAGGCGAGUGAGGUCU AGUGUCGA 1561 722 AACUCCUG G CCAUGGCA 1331 UGCCAUGG GCCGAAAGGCGAGUGAGGUCU CAGGAGUU 1562 728 UGGCCAUG G CACUGGAG 1332 CUCCAGUG GCCGAAAGGCGAGUGAGGUCU CAUGOCCA 1563 736 GCACUGGA G UGCUGGCU 1333 AGCCAGCA GCCGAAAGGCGAGUGAGGUCU UCCAGUGC 1564 738 ACUGGAGU G CUGGCUCU 1334 AGAGCCAG GCCGAAAGGCGAGUGAGGUCU ACUCCAGU 1565 742 GAGUCCUG G CUCUCCCU 1335 AGGGAGAG GCCGAAAGGCGAGUGAGGUCU CACCACUC 1566 754 UCCCUAGG G CACCCUUU 1336 AAAGGGUG GCCGAAAGGCGAGUGAGGUCU CCUAGGGA 1567 775 UACCGACG G CACAUCAC 1337 GUGAUGUG GCCGAAAGGCGAGUGAGGUCU CGUCGGUA 1568 787 AUCACCCU G CGCCUGGG 1338 CCCAGGCG GCCGAAAGGCGAGUGAGGUCU AGGGUGAU 1569 789 CACCCUGC G CCUGGGCG 1339 CGCCCAGG GCCGAAAGGCGAGUGAGGUCU GCAGGGUG 1570 795 GCGCCUGG G CGCACUGG 1340 CCAGUGCG GCCGAAAGGCGAGUGAGGUCU CCAGGCGC 1571 797 GCCUGGGC G CACUGGUG 1341 CACCAGUG GCCGAAAGGCGAGUGAGGUCU GCCCAGGC 1572 803 GCGCACUG G UGGCCCCG 1342 CGGGGCCA GCCGAAAGGCGAGUGAGGUCU CAGUGCGC 1573 806 CACUGGUG G CCCCGGUG 1343 CACCGGGG GCCGAAAGGCGAGUGAGGUCU CACCAGUG 1574 812 UGGCCCCG G UGGUGAGC 1344 GCUCACCA GCCGAAAGGCGAGUGAGGUCU CGGGGCCA 1575 815 CCCCGGUG G UGAGCGCC 1345 GGCGCUCA GCCGAAAGGCGAGUGAGGUCU CACCGGGG 1576 819 GGUGGUGA G CGCCUUCU 1346 AGAAGGCG GCCGAAAGGCGAGUGAGGUCU UCACCACC 1577 821 UGGUGAGC G CCUUCUCC 1347 GGAGAAGG GCCGAAAGGCGAGUGAGGUCU GCUCACCA 1578 833 UCUCCCUG G CUUUCUGC 1348 GCAGAAAG GCCGAAAGGCGAGUGAGGUCU CAGGGAGA 1579 840 GGCUUUCU G CGCGCUAC 1349 GUAGCGCG GCCGAAAGGCGAGUGAGGUCU AGAAAGCC 1580 842 CUUUCUGC G CGCUACCU 1350 AGGUAGCG GCCGAAAGGCGAGUGAGGUCU GCAGAAAG 1581 844 UUCUGCGC G CUACCUUU 1351 AAAGGUAG GCCGAAAGGCGAGUGAGGUCU GCGCAGAA 1582 858 UUUCAUGG G CUUCGGGA 1352 UCCCGAAG GCCGAAAGGCGAGUGAGGUCU CCAUGAAA 1583 868 UUCGGGAA G UUCGUGCA 1353 UGCACGAA GCCGAAAGGCGAGUGAGGUCU UUCCCGAA 1584 872 GGAAGUUC G UCCAGUAC 1354 GUACUGCA GCCGAAAGGCGAGUGAGGUCU GAACUUCC 1585 874 AAGUUCGU G CAGUACUG 1355 CAGUACUG GCCGAAAGGCGAGUGAGGUCU ACGAACUU 1586 877 UUCGUGCA G UACUGCCC 1356 GGGCAGUA CCCGAAAGGCGAGUGAGGUCU UGCACGAA 1587 882 GCAGUACU G CCCCGGCA 1357 UGCCGGGG GCCGAAAGGCCAGUGAGGUCU AGUACUGC 1588 888 CUGCCCCG G CACCUGGU 1358 ACCAGGUG GCCGAAAGGCGAGUGAGGUCU CGGGGCAG 1589 895 GOCACCUG G UGCUUUAU 1359 AUAAAGCA GCCGAAAGGCGAGUGAGGUCU CAGGUGCC 1590 897 CACCUGGU G CUUUAUCC 1360 GGAUAAAG GCCGAAAGGCGAGUGAGGUCU ACCAGGUG 1591 911 UCCAGAUG G UCCACGAG 1361 CUCCUGGA GCCGAAAGGCGAGUGAGGUCU CAUCUGGA 1592 924 CGAGGAGG G CUCGCUGU 1362 ACAGCGAG GCCGAAAGGCGAGUGAGGUCU CCUCCUCG 1593 928 GAGGGCUC G CUGUCGGU 1363 ACCGACAG GCCGAAAGGCGAGUGAGGUCU GAGCCCUC 1594 931 GGCUCGCU G UCGGUGCU 1364 AGCACCGA GCCGAAAGGCGAGUGAGGUCU AGCGAGCC 1595 935 CGCUGUCG G UGCUGGGG 1365 CCCCAGCA GCCGAAAGGCGAGUGAGGUCU CGACAGCG 1596 937 CUGUCGGU G CUGCGGUA 1366 UACCCCAG GCCGAAACCCGAGUGAGGUCU ACCGACAG 1597 943 GUCCUGOG G UACUCUGU 1367 ACAGAGUA GCCGAAAGGCGAGUGAGGUCU CCCAGCAC 1598 950 CGUACUCU G UCCUCUAC 1368 GUACAGCA GCCGAAAGCCGAGUGAGGUCU AGACUACC 1599 952 UACUCUGU G CUCUACUC 1369 GAGUAGAC GCCGAAAGCCGAGUCACGUCU ACAGAGUA 1600 963 CUACUCCA G CCUCAUGG 1370 CCAUGACG CCCGAAACGCGACUGAGGUCU UGGAGUAG 1601 971 GCCUCAUG G CGCUGCUG 1371 CAGCAGCG GCCCAAAGGCGAGUGAGGUCU CAUGAGGC 1602 973 CUCAUGOC G CUGCUCCU 1372 ACCAGCAG GCCGAAAGGCGAGUGAGCUCU OCCAUGAG 1603 976 AUGGCGCU G CUCGUCCU 1373 AGGACCAG GCCGAAAGGCGAGUCAGGUCU AGCGCCAU 1604 980 CGCUGCUG G UCCUCGCC 1374 GGCGACGA GCCGAAAGGCGACUGAGGUCU CAGCAGCG 1605 986 UGGUCCUC G CCACCGUC 1375 CACGGUGG GCCGAAAGGCGAGUGACGUCU GAGGACCA 1606 992 UCCCCACC G UGCUGUGC 1376 GCACAGCA GCCGAAAGGCGAGUGAGGUCU GGUGGCGA 1607 994 GCCACCGU G CUGUGCAA 1377 UUGCACAG GCCGAAAGGCGAGUGACGUCU ACGGUGGC 1608 997 ACCCUGCU G UGCAACCU 1378 AGCUUGCA GCCGAAAGGCGACUGAGGUCU AGCACCCU 1609 999 CGUGCUGU G CAACCUCG 1379 CGAGGUUG GCCCAAAGGCGAGUGAGGUCU ACAGCACG 1610 1008 CAACCUCC G CCCCAUGC 1380 GCAUGGCG GCCGAAAGGCCAGUCAGGUCU CGACGUUG 1611 1010 ACCUCGGC G CCAUGCGC 1381 GCGCAUGG GCCGAAAGGCGAGUGAGGUCU CCCGAGGU 1612 1015 GGCCCCAU G CCCAACCU 1382 AGGUUGCC GCCGAAAGGCCAGUGAGGUCU AUGCCCCC 1613 1017 CGCCAUCC G CAACCUCU 1383 AGAGGUUG CCCGAPAGGCGACUGAGGUCU GCAUGGCG 1614 1028 ACCUCUAU G CGAUGCAC 1384 GUCCAUCG GCCCAAAGGCGAGUGAGCUCU AUAGAGGU 1615 1033 UAUGCGAU G CACCGCCC 1385 CGCCGGUG GCCCAAACGCGAGUGAGGUCU AUCGCAUA 1616 1039 AUGCACCC G CUCCUCCA 1386 UCCAGCCC CCCCAAAGGCGAGUCACCUCU CGGUCCAU 1617 1042 CACCCGCC G CUCCACCC 1387 CCCUGCAG CCCGAAACCCCACUGAGGUCU CCCCCCUG 1618 1045 CGGCCCCU G CACCUCCA 1388 UCCCGCUG CCCCAAAGGCGAGUCACCUCU AGCCGCCC 1619 1048 CCCCUGCA G CCCCACCC 1389 CCCUCCCC GCCGAAAGCCCACUGACGUCU UGCACCCG 1620 1051 CUCCACCC G CACCCCCG 1390 OCCUGGUC CCCCAAAGGCGAGUCAGCUCU CGCUGCAG 1621 1057 CCCCACCC G CUCUCCUC 1391 CACCACCG GCCGAAAGCCCAGUGAGCUCU CCCUCCCG 1622 1059 CCACCCCC G CUCCUCCA 1392 UCCACCAC CCCCAAAGGCGACUCACCUCU GCGGCUCC 1623 1065 GCCCUCCU G CACCACGG 1393 CCCUGGUG GCCCAAACCCCAGUCAGCUCU ACCACCUC 1624 1077 CACGGACU G UCCCCACC 1394 GCUCCCCA GCCGAAACCCGACUCAGGUCU AGUCCCUC 1625 1079 CCCACUCU G CCCAGCCC 1395 CUCCUCUC CCCCAAAGCCGAGUGACCUCU ACAGUCCC 1626 1084 UCUGCCCA G CCCCCCGC 1396 GCGCGCGG CCCCAAACCCCAGUGACGUCU UCCCCACA 1627 1087 CCCCAGCC G CUCUCUCA 1397 UCCCCCCC GCCGAAAGGCGACUCACGUCU GUCUCCUC 1628 1089 CGAGCCGC G CGCGGACG 1398 CGUCCGCG CCCCAAACCCCAGUGAGGUCU CCCGCUCG 1629 1091 AUCCUCUC G CGGACCCC 1399 CCCCUCCG GCCGAAAGGCGACUCACGUCU CCGCCCCU 1630 1106 CCAGGCAA G CCUCCCCU 1400 AGGGGACG CCCGAAACCCGAGUGAGCUCU UUCCCUCC 1631 1108 ACCCAAGC G UCCCCUCA 1401 UCAGCCCA GCCGAAAGGCCAGUCACCUCU CCUUCCCU 1632 1117 UCCCCUCA G CCCCUCGA 1402 UCCAGGUC CCCGAAACCCGAGUCAGCUCU UCAGUGGA 1633 1129 CUGGAGGA G CUCCAUCA 1403 UCAUCCAG GCCGAAACCCCACUCAGGUCU UCCUCCAC 1634 1144 CACCUCCU G CUCCUGUC 1404 UCCACCAC CCCCAAAGGCGACUGACCUCU AGGAGGUC 1635 1147 CUCCUCCU G CUGCCGCU 1405 ACCGCCAG CCCGAAACCCGAGUGAGCUCU ACCACCAG 1636 1151 UCCUCCUC G CUCUCAUG 1406 CAUCACCC GCCGAAAGCCGACUGAGGUCU CACCACCA 1637 1153 CUGCUGGC G CUGAUGAC 1407 GUCAUCAG GCCGAAAGGCGAGUGAGGUCU UCCACCAG 1638 1163 UGAUGACC G UGCUCUUC 1408 GAAGAGCA GCCGAAAGGCGAGUCAGGUCU GGUCAUCA 1639 1165 AUGACCGU G CUCUUCAC 1409 GUGAAGAG GCCGAAAGGCGAGUGAGGUCU ACGGUCAU 1640 1177 UUCACUAU G UGUUCUCU 1410 AGAGAACA GCCGAAACCCGAGUGAGGUCU AUAGUGAA 1641 1179 CACUAUGU G UUCUCUGC 1411 GCAGAGAA GCCGAAAGGCGACUGACCUCU ACAUACUC 1642 1186 UGUUCUCU G CCCGUAAU 1412 AUUACGGG GCCGAAAGGCGAGUGAGGUCU AGAGAACA 1643 1190 CUCUGCCC G UAAUUUAU 1413 AUAAAUUA GCCCAAAGGCGAGUCAGGUCU GGGCAGAG 1644 1200 AAUUUAUC G CGCUUACU 1414 AGUAAGCG GCCGAAAGGCGACUCAGGUCU GAUAAAUU 1645 1202 UUUAUCGC G CUUACUAU 1415 AUAGUAAG GCCCAAACCCCAGUGAGGUCU GCCAUAAA 1646 1214 ACUAUGCA G CAUUUAAC 1416 CUUAAAUG CCCGAAAGGCGAGUGACCUCU UCCAUACU 1647 1226 UUAAGGAU G UCAACCAG 1417 CUCCUCCA GCCGAAACGCGAGUGAGCUCU AUCCUUAA 1648 1256 CUGAAGAA G CAGAAGAC 1418 GUCUUCUG CCCCAAACCCGACUGAGCUCU UUCUUCAG 1649 1271 ACCUCCGA G CCUUGCCA 1419 UCGCAAGG GCCGAAACGCGAGUGAGCUCU UCGGAGGU 1650 1276 CCAGCCUU G CCAUUUCU 1420 AGAAAUCG GCCGAAACCCGACUGAGGUCU AACGCUCG 1651 1289 UUCUAUCU G UGAUCUCA 1421 UCAAAUCA CCCGAAAGCCCAGUGAGGUCU AGAUAGAA 1652 1301 UUUCAAUU G UGGACCCU 1422 ACCGUCCA GCCCAAACGCCAGUGAGGUCU AAUUCAAA 1653 1337 GAUCUCCA G UAUUUCCG 1423 CCGAAAUA CCCGAAAGGCGAGUGAGGUCU UGGAGAUC 1654 1381 CCUCUUAG G UACAGGAG 1424 CUCCUGUA GCCGAAAGGCGAGUGAGCUCU CUAAGAGG 1655 1389 CUACAGGA G CCGGUGCA 1425 UGCACCGC GCCGAAACCCGAGUGAGGUCU UCCUGUAC 1656 1393 AGGAGCCG G UCCAGCAA 1426 UUCCUGCA CCCCAAAGGCGAGUCACCUCU CGGCUCCU 1657 1395 CACCCCCU G CAGCAAUU 1427 AAUUCCUC GCCGAAACCCCACUGAGGUCU ACCCCCUC 1658 1398 CCGCUGCA G CAAUUCCA 1428 UGGAAUUG CCCCAAAGGCCAGUCACCUCU UGCACCCG 1659 1422 CCAAUCCA G UCUGUCAC 1429 CUCACACA CCCCAAACCCCACUGAGGUCU UCCAUUCC 1660 1426 UCCAGUCU G UCACACUC 1430 CACUGUCA CCCCAAAGGCCAGUCACCUCU AGACUGGA 1661 1432 CUCUCACA G UCUUUUUC 1431 CAAAAACA CCCGAAACCCCACUGAGCUCU UCUCACAG 1662 1434 CUGACACU G UUUUUCAC 1432 CUGAAAAA CCCGAAACGCCACUCACCUCU ACUGUCAC 1663 1446 UUCACUCU G UCCUAAGC 1433 CCUUACCA CCCCAAACGCGAGUGACCUCU AGACUGAA 1664 1449 ACUCUGUG G UAACCUCA 1434 UCACCUUA GCCGAAACCCCACUGAGCUCU CACACACU 1665 1453 UCUCCUAA G CUGAGGAA 1435 UUCCUCAC CCCGAAAGGCGACUCACCUCU UUACCACA 1666 1465 ACCAAUAU G UCACAUUU 1436 AAAUCUGA CCCCAAACCCGAGUGACCUCU AUAUUCCU 1667 1477 CAUUUUCA G UCAAACAA 1437 UUCUUUCA CCCCAAACCCCACUCACCUCU UGAAAAUC 1668

[0233] TABLE VI Human PTGDR DNAzyme and Substrate Sequence Seq Seq Pos Substrate ID DNAzyme ID 9 GAAUUCUG G CUAUUUUC 1207 GAAAATAG GGCTAGCTACAACGA CAGAATTC 1715 12 UUCUGGCU A UUUUCCUC 1 GAGGAAAA GGCTAGCTACAACGA AGCCAGAA 1716 23 UUCCUCCU G CCGUUCCG 1208 CGGAACGG GGCTAGCTACAACGA AGGAGGAA 1717 26 CUCCUGCC G UUCCGACU 1209 AGTCGGAA GGCTAGCTACAACGA GGCAGGAG 1718 32 CCGUUCCG A CUCGGCAC 1669 GTGCCGAG GGCTAGCTACAACGA CGGAACGG 1719 37 CCGACUCG G CACCAGAG 1210 CTCTGGTG GGCTAGCTACAACGA CGAGTCGG 1720 39 GACUCGGC A CCAGAGUC 463 GACTCTGG GGCTAGCTACAACGA GCCGAGTC 1721 45 GCACCAGA G UCUGUCUC 1211 GAGACAGA GGCTAGCTACAACGA TCTGGTGC 1722 49 CAGAGUCU G UCUCUACU 1212 AGTAGAGA GGCTACCTACAACGA AGACTCTG 1723 55 CUGUCUCU A CUGAGAAC 13 GTTCTCAG GGCTAGCTACAACGA AGAGACAG 1724 62 UACUGAGA A CGCAGCGC 1670 GCGCTGCG GGCTAGCTACAACGA TCTCAGTA 1725 64 CUGAGAAC G CAGCGCGU 1213 ACGCGCTG GGCTAGCTACAACGA GTTCTCAG 1726 67 AGAACGCA G CGCGUCAG 1214 CTGACGCG GGCTAGCTACAACGA TGCGTTCT 1727 69 AACGCAGC G CGUCAGGG 1215 CCCTGACG GGCTAGCTACAACGA GCTGCGTT 1728 71 CGCAGCGC G UCAGGGCC 1216 GGCCCTGA GGCTAGCTACAACGA GCGCTGCG 1729 77 GCGUCAGG G CCGAGCUC 1217 CAGCTCGG GGCTAGCTACAACCA CCTGACGC 1730 82 AGCCCCGA G CUCUUCAC 1218 GTCAAGAC GCCTAGCTACAACGA TCGGCCCT 1731 89 AGCUCUUC A CUGGCCUG 475 CAGGCCAG GGCTAGCTACAACGA GAAGAGCT 1732 93 CUUCACUG G CCUGCUCC 1219 GCACCAGG GGCTAGCTACAACGA CAGTGAAG 1733 97 ACUCCCCU G CUCCGCGC 1220 GCGCGGAG GGCTAGCTACAACGA AGOCCACT 1734 102 CCUGCUCC G CCCUCUUC 1221 CAAGAGCG GGCTAGCTACAACGA GGACCAGG 1735 104 UGCUCCGC G CUCUUCAA 1222 TTCAAGAG GGCTAGCTACAACGA GCGGAGCA 1736 112 GCUCUUCA A UGCCAGCG 1671 CGCTGGCA GGCTAGCTACAACGA TGAAGAGC 1737 114 UCUUCAAU G CCAGCGCC 1223 GGCGCTGG GGCTAGCTACAACGA ATTGAAGA 1738 118 CAAUGCCA G CGCCAGGC 1224 GCCTGGCG GGCTAGCTACAACGA TGGCATTG 1739 120 AUGCCAGC G CCAGCCGC 1225 GCGCCTGG GGCTAGCTACAACGA GCTGCCAT 1740 125 AGCGCCAG G CGCUCACC 1226 GGTGAGCG CGCTAGCTACAACGA CTGGCGCT 1741 127 CGCCAGGC G CUCACCCU 1227 AGGGTGAG GGCTAGCTACAACGA GCCTGGCG 1742 131 AGGCGCUC A CCCUGCAG 489 CTGCAGGG GGCTAGCTACAACGA GAGCGCCT 1743 136 CUCACCCU G CAGAGCGU 1228 ACGCTCTG GGCTAGCTACAACGA AGGGTGAG 1744 141 CCUGCAGA G CGUCCCGC 1229 GCGGGACG GGCTAGCTACAACGA TCTGCAGG 1745 143 UGCAGAGC G UCCCGCCU 1230 AGGCGGGA GGCTAGCTACAACGA GCTCTGCA 1746 148 AGCGUCCC G CCUCUCAA 1231 TTGAGAGG GGCTAGCTACAACGA GGGACGCT 1747 163 AAAGAGGG G UGUGACCC 1232 GGGTCACA GGCTAGCTACAACGA CCCTCTTT 1748 165 AGAGGGGU G UGACCCGC 1233 GCGGGTCA GGCTAGCTACAACGA ACCCCTCT 1749 168 GCGGUGUG A CCCGCGAG 1672 CTCGCGGG GGCTAGCTACAACGA CACACCCC 1750 172 UGUGACCC G CGAGUUUA 1234 TAAACTCG GGCTAGCTACAACGA GGGTCACA 1751 176 ACCCGCGA G UUUAGAUA 1235 TATCTAAA GGCTAGCTACAACGA TCGCGGGT 1752 182 GAGUUUAG A UAGGAGGU 1673 ACCTCCTA GGCTAGCTACAACGA CTAAACTC 1753 189 GAUAGGAG G UUCCUGCC 1236 GGCAGGAA GGCTAGCTACAACGA CTCCTATC 1754 195 AGGUUCCU G CCGUGGGG 1237 CCCCACCG GGCTAGCTACAACGA AGGAACCT 1755 198 UUCCUGCC G UGGGGAAC 1238 GTTCCCCA GGCTAGCTACAACGA GCCAGGAA 1756 205 CGUGGGGA A CACCCCGC 1674 CCCGGGTG GGCTAGCTACAACGA TCCCCACG 1757 207 UGGCGAAC A CCCCGCCG 505 CGGCGGGG GGCTAGCTACAACGA GTTCCCCA 1758 212 AACACCCC G CCCCCCUC 1239 GAGCCCCG GGCTAGCTACAACGA GCGGTGTT 1759 215 ACCCCGCC G CCCUCGGA 1240 TCCGAGCG GGCTAGCTACAACGA GGCGGCGT 1760 224 CCCUCGGA G CUUUUUCU 1241 AGAAAAAG GGCTAGCTACAACGA TCCGAGGG 1761 233 CUUUUUCU G UGGCGCAG 1242 CTGCGCCA GGCTAGCTACAACGA AGAAAAAG 1762 236 UUUCUCUG G CGCAGCUU 1243 AAGCTGCC GGCTAGCTACAACGA CACAGAAA 1763 238 UCUGUGGC G CAGCUUCU 1244 AGAAGCTG GGCTAGCTACAACGA GCCACAGA 1764 241 GUGGCGCA G CUUCUCCG 1245 CGGAGAAG GCCTAGCTACAACGA TOCCCCAC 1765 249 GCUUCUCC G CCCGAGCC 1246 GGCTCGGG CGCTAGCTACAACGA GGAGAAGC 1766 255 CCGCCCGA G CCCCGCGC 1247 GCGCGCGG GGCTACCTACAACGA TCGGGCGG 1767 258 CCCGAGCC G CGCGCGGA 1248 TCCGCGCG GGCTAGCTACAACCA GGCTCCCG 1768 260 CGAGCCGC G CGCGGAGC 1249 GCTCCCCG GGCTAGCTACAACGA GCGGCTCG 1769 262 ACCCGCCC G CGGAGCUG 1250 CAGCTCCG GGCTAGCTACAACGA CCGCGCCT 1770 267 CGCGCGGA G CUCCCGCG 1251 CCCGGCAG GGCTAGCTACAACCA TCCGCGCG 1771 270 GCGCACCU G CCGGCGGC 1252 GCCCCCCG GGCTAGCTACAACGA AGCTCCGC 1772 277 UGCCGGGG G CUCCUUAG 1253 CTAAGGAC GGCTAGCTACAACCA CCCCGGCA 1773 285 CCUCCUUA G CACCCCGG 1254 CCCCGGTC GCCTAGCTACAACGA TAAGGAGC 1774 287 UCCUUAGC A CCCCGGCG 527 CCCCCGGG GGCTAGCTACAACGA CCTAAGGA 1775 293 GCACCCGG G CGCCGCCG 1255 CCCCGGCG CCCTAGCTACAACCA CCGGCTGC 1776 295 ACCCCCCC G CCGGGCCC 1256 CGCCCCGC GCCTAGCTACAACGA CCCCGGCT 1777 301 CCGCCGGC G CCCUCGCC 1257 GGCGAGGG GGCTAGCTACAACGA CCCCCCCC 1778 307 GGGCCCUC G CCCUUCCG 1258 CGCAAGCC GCCTAGCTACAACCA GAGGGCCC 1779 315 GCCCUUCC G CAGCCUUC 1259 GAAGGCTG GCCTAGCTACAACGA GGAACGGC 1780 318 CUUCCGCA G CCUUCACU 1260 ACTCAAGC GGCTACCTACAACCA TGCCGAAG 1781 324 CAGCCUUC A CUCCACCC 541 CCCTCGAG GGCTACCTACAACGA CAACCCTG 1782 330 UCACUCCA G CCCUCUGC 1261 GCACACGG GGCTAGCTACAACCA TGGACTCA 1783 337 ACCCCUCU G CUCCCGCA 1262 TCCGCGAG GCCTACCTACAACGA ACAGGCCT 1784 343 CUGCUCCC G CACCCCAU 1263 ATGCCGTG CCCTACCTACAACGA CCGACCAC 1785 345 GCUCCCCC A CGCCAUCA 552 TCATGCCC CGCTACCTACAACCA GCGGCAGC 1786 347 UCCCGCAC G CCAUCAAG 1264 CTTCATGC GCCTAGCTACAACCA GTGCGGGA 1787 350 CGCACGCC A UCAACUCC 554 CGACTTCA GGCTACCTACAACGA GGCGTCCG 1788 355 CCCAUCAA G UCGCCGUU 1265 AACGCCGA GGCTAGCTACAACCA TTCATCGC 1789 358 AUCAACUC G CCCUUCUA 1266 TACAACCG CGCTAGCTACAACGA GACTTCAT 1790 361 AAGUCGCC G UUCUACCC 1267 CGCTACAA GGCTAGCTACAACCA CCCGACTT 1791 366 GCCCUUCU A CCCCUGCC 55 GCCAGCGG GCCTAGCTACAACCA ACAACCGC 1792 369 GUUCUACC G CUGCCAGA 1268 TCTGCCAG GCCTACCTACAACGA GGTACAAC 1793 372 CUACCCCU G CCAGAACA 1269 TGTTCTCC GCCTAGCTACAACCA AGCCGTAG 1794 378 CUCCCACA A CACCACCU 1675 ACCTCCTC GCCTACCTACAACGA TCTCCCAC 1795 380 GCCACAAC A CCACCUCU 561 ACACCTCC CCCTACCTACAACCA CTTCTCCC 1796 383 AGAACACC A CCUCUCUG 563 CACAGAGG CCCTACCTACAACGA CCTCTTCT 1797 389 CCACCUCU G UCGAAAAA 1270 TTTTTCCA CGCTAGCTACAACCA AGAGCTCC 1798 399 CCAAAAAG G CAACUCCG 1271 CCCAGTTG CCCTACCTACAACGA CTTTTTCC 1799 402 AAAACCCA A CUCGCCCC 1676 CCCCCCAC CCCTACCTACAACCA TGCCTTTT 1800 407 CCAACUCG G CCCUCAUC 1272 CATCACCC GCCTACCTACAACCA CCACTTCC 1801 410 ACUCCCCC G UGAUCGCC 1273 CCCCATCA CCCTACCTACAACQA CGCCGACT 1802 413 CCGCCCUG A UCCCCCCC 1677 CCCCCCCA CCCTACCTACAACCA CACCCCCC 1803 417 CCUCAUCC G CCGCCUCC 1274 CCACCCCC CGCTAGCTACAACCA CCATCACC 1804 422 UCCCCCCC G UCCUCUUC 1275 CAACACCA CCCTACCTACAACCA CCCCCCCA 1805 424 CCCCCGCU G CUCUUCAG 1276 CTCAACAC CCCTACCTACAACGA ACCCCCCC 1806 432 CCUCUUCA G CACCCCCC 1277 CCCCCCTC CCCTACCTACAACCA TCAAGACC 1807 434 UCUUCACC A CCGGCCUC 572 CACGCCCC CCCTACCTACAACCA CCTCAAGA 1808 438 CACCACCG G CCUCCUCC 1278 CCACCAGC GCCTACCTACAACGA CGCTCCTC 1809 447 CCUCCUCC G CAACCUGC 1279 CCACCTTC CCCTAGCTACAACCA CCAGGAGC 1810 450 CCUGCCCA A CCUCCUCC 1678 CCACCAGC GCCTACCTACAACGA TCCCCACC 1811 454 GCCAACCU G CUGGCCCU 1280 AGGGCCAC GGCTAGCTACAACGA AGGTTGCC 1812 458 ACCUGCUG G CCCUGGGG 1281 CCCCAGGG GGCTAGCTACAACGA CAGCAGGT 1813 466 GCCCUGGG G CUGCUCGC 1282 GCCAGCAG GGCTAGCTACAACGA CCCAGCGC 1814 469 CUCGGGCU G CUGGCGCG 1283 CGCGCCAC CGCTAGCTACAACGA AGCCCCAG 1815 473 GCCUGCUG G CCCGCUCG 1284 CGAGCGCG GGCTAGCTACAACGA CAGCAGCC 1816 475 CUGCUGGC G CGCUCGGG 1285 CCCGAGCG CGCTAGCTACAACGA GCCAGCAG 1817 477 GCUCGCGC G CUCCCGGC 1286 GCCCCGAC GGCTACCTACAACGA GCGCCAGC 1818 484 CCCUCGGG G CUGGCGUG 1287 CACCCCAG GGCTAGCTACAACCA CCCGAGCG 1819 490 GGCCUGGC G UGCUGCUC 1288 CACCACCA GCCTACCTACAACGA CCCAGCCC 1820 493 CUCCGGUG G UGCUCCCC 1289 CGCCAGCA CCCTAGCTACAACGA CACCCCAG 1821 495 CCGGUCGU G CUCCCGGC 1290 CCCCCGAG GGCTAGCTACAACCA ACCACCCC 1822 499 UCGUGCUC G CCGCCUCC 1291 CCACCCCC GGCTAGCTACAACGA CAGCACCA 1823 502 UCCUCGCC G CCUCCACU 1292 ACTGGACG GCCTACCTACAACCA CGCCAGCA 1824 504 CUCGCGCC G UCCACUCC 1293 CCAGTCGA GGCTAGCTACAACCA GCCGCGAG 1825 508 CCCCGUCC A CUCCCCCC 591 GCGCGCAG GCCTAGCTACAACGA CGACGCCC 1826 511 CGUCCACU G CGCCCGCU 1294 AGCGGGCG GGCTAGCTACAACGA AGTCCACC 1827 513 UCCACUCC G CCCGCUGC 1295 GCACCCGC CGCTAGCTACAACCA GCACTCGA 1828 517 CUGCGCCC G CUCCCCUC 1296 CAGGUCAG GGCTACCTACAACGA GGGCGCAG 1829 520 CCCCCCCU G CCCUCCGU 1297 ACCGAGGG GGCTAGCTACAACCA AGCCCCCG 1830 527 UGCCCUCC G UCUUCUAC 1298 GTAGAAGA GCCTAGCTACAACGA CCAGGGCA 1831 534 GCUCUUCU A CAUGCUCC 69 CCACCATG GGCTAGCTACAACCA ACAAGACC 1832 536 UCUCCUAC A UGCUGCUC 601 CACCACCA CCCTACCTACAACGA GTAGAAGA 1833 538 UUCUACAU G CUGGUGUG 1299 CACACCAG CGCTAGCTACAACGA ATGTAGAA 1834 542 ACAUGCUG G UCUCUCCC 1300 CCCACACA CGCTACCTACAACCA CACCATGT 1835 544 AUGCUGGU G UGUGGCCU 1301 ACGCCACA GCCTACCTACAACGA ACCAGCAT 1836 546 CCUGGUGU G UCCCCUGA 1302 TCACGCCA GGCTAGCTACAACCA ACACCAGC 1837 549 GGUCUGUG G CCUGACGG 1303 CCGTCAGC GCCTAGCTACAACGA CACACACC 1838 554 CUGCCCUG A CCGUCACC 1679 GGTGACCC GGCTAGCTACAACGA CAGGOCAC 1839 557 CCCUCACC G UCACOGAC 1304 CTCCCTGA CCCTACCTACAACGA CCTCACCC 1840 560 UCACCCUC A CCCACUUC 605 CAACTCCC CGCTAGCTACAACCA GACCGTCA 1841 564 GGUCACCC A CUUCCUGG 1680 CCACCAAC CCCTACCTACAACCA CCCTCACC 1842 568 ACCGACUU G CUCCCCAA 1305 TTCCCCAC GCCTAGCTACAACCA AAGTCGCT 1843 573 CUUCCUCC G CAACUCCC 1306 GGCACTTC CCCTAGCTACAACCA CCACCAAG 1844 577 CUCCCCAA G UCCCUCCU 1307 ACGACGCA CCCTACCTACAACGA TTCCCCAC 1845 579 CCGCAAGU G CCUCCUAA 1308 TTACCACG CCCTACCTACAACCA ACTTGCCC 1846 588 CCUCCUAA G CCCGGUCC 1309 CCACCCCC CCCTAGCTACAACCA TTACCAGG 1847 593 UAAGCCCC G UCCUCCUG 1310 CACCACCA GGCTACCTACAACGA CGGGCTTA 1848 596 CCCCCCUG G UGCUGCCU 1311 AGCCACCA CCCTAGCTACAACCA CACCCGGC 1849 598 CCGGUGGU G CUCCCUGC 1312 CCACCCAG GGCTACCTACAACGA ACCACCCC 1850 602 UCCUCCUC G CUGCCUAC 1313 GTAGCCAC CCCTAGCTACAACCA CACCACCA 1851 605 UGCUGGCU G CCUACCCU 1314 ACCCTAGG GGCTACCTACAACGA AGCCACCA 1852 609 CCCUCCCU A CGCUCACA 74 TCTCACCC CCCTAGCTACAACCA ACCCACCC 1853 611 CUGCCUAC G CUCAGAAC 1315 CTTCTGAG CCCTACCTACAACGA GTACCCAG 1854 618 CGCUCAGA A CCCCACUC 1681 CACTCCCG CCCTAGCTACAACCA TCTGAGCC 1855 624 CAACCCGA G UCUCCCCC 1316 CCCCCACA CCCTACCTACAACCA TCCCCTTC 1856 628 CCCACUCU G CGCGUGCU 1317 ACCACCCG CCCTACCTACAACGA AGACTCCC 1857 632 GUCUGCGC G UCCUUCCG 1318 CCCAACCA CGCTAGCTACAACCA CCCCACAC 1858 634 CUCCCCCU G CUUGCCCC 1319 GGCGCAAG CCCTAGCTACAACGA ACCCCCAG 1859 638 CGGUGCUU G CCCCCCCA 1320 TCCCCCCG GGCTACCTACAACCA AACCACCC 1860 640 CUCCUUCC G CCCGCAUU 1321 AATCCGGG CCCTAGCTACAACGA CCAACCAC 1861 644 UUGCCCCC G CAUUCCAC 1322 GTCCAATC GCCTACCTACAACCA GGCCGCAA 1862 646 GCGCCCGC A UUGGACAA 627 TTGTCCAA GGCTAGCTACAACGA GCGGGCGC 1863 651 CGCAUUGG A CAACUCGU 1682 ACGAGTTG GGCTAGCTACAACGA CCAATGCG 1864 654 AUUGGACA A CUCGUUGU 1683 ACAACGAG GGCTAGCTACAACGA TGTCCAAT 1865 658 GACAACUC G UUGUGCCA 1323 TGGCACAA GGCTAGCTACAACGA GAGTTGTC 1866 661 AACUCGUU G UGCCAAGC 1324 GCTTGGCA GGCTAGCTACAACGA AACGAGTT 1867 663 CUCGUUGU G CCAAGCCU 1325 AGGCTTGG GGCTAGCTACAACGA ACAACGAG 1868 668 UGUGCCAA G CCUUCGCC 1326 GGCGAAGG GGCTAGCTACAACGA TTGGCACA 1869 674 AAGCCUUC G CCUUCUUC 1327 GAAGAAGG GGCTAGCTACAACGA GAAGGCTT 1870 683 CCUUCUUC A UGUCCUUC 637 GAAGGACA GGCTAGCTACAACGA GAAGAAGG 1871 685 UUCUUCAU G UCCUUCUU 1328 AAGAAGGA CGCTAGCTACAACGA ATGAAGAA 1872 697 UUCUUUGG G CUCUCCUC 1329 GAGGAGAG GGCTAGCTACAACGA CCAAAGAA 1873 707 UCUCCUCG A CACUGCAA 1684 TTGCAGTG GGCTAGCTACAACGA CUAGGAGA 1874 709 UCCUCGAC A CUGCAACU 645 AGTTGCAG GGCTAGCTACAACGA GTCGAGGA 1875 712 UCGACACU G CAACUCCU 1330 AGGAGTTG GGCTAGCTACAACGA AGTGTCGA 1876 715 ACACUGCA A CUCCUGGC 1685 GCCAGGAG GGCTAGCTACAACGA TGCAGTGT 1877 722 AACUCCUG G CCAUGGCA 1331 TGCCATGG GGCTAGCTACAACGA CAGGAGTT 1878 725 UCCUGGCC A UGGCACUG 652 CAGTGCCA GGCTAGCTACAACGA GGCCAGGA 1879 728 UGGCCAUG G CACUGGAG 1332 CTCCAGTG GGCTAGCTACAACGA CATGGCCA 1880 730 GCCAUGGC A CUGGAGUG 653 CACTCCAG GGCTAGCTACAACGA GCCATGGC 1881 736 GCACUGGA G UGCUGGCU 1333 AGCCAGCA GGCTAGCTACAACGA TCCAGTGC 1882 738 ACUGGAGU G CUGGCUCU 1334 AGAGCCAG GGCTAGCTACAACGA ACTCCAGT 1883 742 GAGUCCUG G CUCUCCCU 1335 AGGGAGAG GCCTAGCTACAACGA CAGCACTC 1884 754 UCCCUAGG G CACCCUUU 1336 AAAGGGTG GGCTAGCTACAACGA CCTAGGGA 1885 756 CCUAGGGC A CCCUUUCU 661 AGAAAGGG GGCTAGCTACAACGA GCCCTAGG 1886 768 UUUCUUCU A CCGACGGC 104 GCCGTCGG GGCTAGCTACAACGA AGAACAAA 1887 772 UUCUACCG A CGGCACAU 1686 ATGTGCCG GGCTAGCTACAACGA CGGTAGAA 1888 775 UACCGACG G CACAUCAC 1337 GTGATGTG GGCTAGCTACAACGA CGTCGGTA 1889 777 CCGACGGC A CAUCACCC 668 GGGTGATG GGCTAGCTACAACGA GCCGTCGG 1890 779 GACGGCAC A UCACCCUG 669 CAGGGTGA GGCTAGCTACAACGA GTCCCGTC 1891 782 CGCACAUC A CCCUGCGC 670 GCGCAGGG GGCTAGCTACAACGA GATGTGCC 1892 787 AUCACCCU G CGCCUGGG 1338 CCCAGGCG GCCTAGCTACAACGA AGGGTGAT 1893 789 CACCCUGC G CCUGGGCG 1339 CGCCCAGG GGCTACCTACAACGA GCAGCGTG 1894 795 GCGCCUGG G CGCACUGG 1340 CCAGTCCG GGCTAGCTACAACGA CCAGGCGC 1895 797 GCCUCGGC G CACUGGUG 1341 CACCAGTG GCCTAGCTACAACGA GCCCAGGC 1896 799 CUGGGCGC A CUGGUGGC 676 GCCACCAG GGCTAGCTACAACGA GCGCCCAG 1897 803 CCCCACUG G UGGCCCCC 1342 CGGGGCCA CGCTAGCTACAACGA CAGTGCGC 1898 806 CACUGGUG G CCCCGGUG 1343 CACCGGGG GGCTAGCTACAACGA CACCAGTG 1899 812 UCGCCCCG G UGGUGAGC 1344 GCTCACCA GGCTAGCTACAACGA CGGGGCCA 1900 815 CCCCGGUG G UGAGCGCC 1345 GGCGCTCA GGCTAGCTACAACGA CACCGGGC 1901 819 GGUGGUGA G CGCCUUCU 1346 AGAAGGCG CGCTAGCTACAACGA TCACCACC 1902 821 UGGUGAGC G CCUUCUCC 1347 GGAGAAGG GGCTAGCTACAACGA GCTCACCA 1903 833 UCUCCCUG G CUUUCUGC 1348 GCACAAAG GGCTAGCTACAACCA CAGGGAGA 1904 840 GGCUUUCU G CGCGCUAC 1349 GTAGCGCG CGCTAGCTACAACGA AGAAAGCC 1905 842 CUUUCUGC G CCCUACCU 1350 ACCTACCG GGCTAGCTACAACGA GCAGAAAG 1906 844 UUCUCCGC G CUACCUUU 1351 AAAGGTAG GCCTAGCTACAACCA CCGCAGAA 1907 847 UGCCCCCU A CCUUUCAU 112 ATGAAAGG GGCTACCTACAACGA AGCGCCCA 1908 854 UACCUUUC A UGCCCUUC 692 CAACCCCA GGCTAGCTACAACCA CAAAGGTA 1909 858 UUUCAUCG G CUUCGGGA 1352 TCCCGAAG GGCTAGCTACAACGA CCATGAAA 1910 868 UUCGGGAA G UUCCUGCA 1353 TGCACGAA GGCTAGCTACAACGA TTCCCGAA 1911 872 GGAAGUUC G UGCAGUAC 1354 GTACTGCA GGCTAGCTACAACGA GAACTTCC 1912 874 AAGUUCGU G CACUACUG 1355 CAGTACTG GGCTAGCTACAACGA ACGAACTT 1913 877 UUCGUGCA G UACUGCCC 1356 GGGCAGTA GGCTAGCTACAACGA TGCACGAA 1914 879 CGUGCAGU A CUGCCCCG 120 CGGGGCAG GGCTAGCTACAACGA ACTGCACG 1915 882 GCAGUACU G CCCCGGCA 1357 TGCCGGGG GGCTAGCTACAACGA AGTACTGC 1916 888 CUGCCCCG G CACCUGGU 1358 ACCAGGTG GGCTAGCTACAACGA CGGGGCAG 1917 890 GCCCCGGC A CCUGGUGC 699 GCACCAGG GGCTAGCTACAACGA GCCGGGGC 1918 895 GGCACCUG G UGCUUUAU 1359 ATAAAGCA GGCTAGCTACAACGA CAGGTGCC 1919 897 CACCUGGU G CUUUAUCC 1360 GGATAAAG GGCTAGCTACAACGA ACCAGGTG 1920 902 GGUGCUUU A UCCAGAUG 123 CATCTGGA GGCTAGCTACAACGA AAAGCACC 1921 908 UUAUCCAG A UGGUCCAC 1687 GTGGACCA GGCTAGCTACAACGA CTGGATAA 1922 911 UCCAGAUG G UCCACGAG 1361 CTCGTGGA GGCTAGCTACAACGA CATCTGGA 1923 915 GAUGGUCC A CGAGGAGG 706 CCTCCTCG GGCTAGCTACAACGA GGACCATC 1924 924 CGAGGAGG G CUCGCUGU 1362 ACAGCGAG GGCTAGCTACAACGA CCTCCTCG 1925 928 GAGGGCUC G CUGUCGGU 1363 ACCUACAG GGCTAGCTACAACGA GAGCCCTC 1926 931 GGCUCGCU G UCGGUGCU 1364 AGCACCGA GGCTAGCTACAACGA AGCGAGCC 1927 935 CGCUGUCG G UGCUGGGG 1365 CCCCAGCA GGCTAGCTACAACGA CGACAGCG 1928 937 CUGUCGGU G CUGGGGUA 1366 TACCCCAG GGCTAGCTACAACGA ACCGACAG 1929 943 GUGCUGGG G UACUCUGU 1367 ACAGAUTA GGCTAGCTACAACGA CCCAGCAC 1930 945 GCUGGGGU A CUCUGUGC 128 GCACAGAG GGCTAGCTACAACGA ACCCCAGC 1931 950 GGUACUCU G UGCUCUAC 1368 GTAGAGCA GGCTAGCTACAACGA AGAGTACC 1932 952 UACUCUGU G CUCUACUC 1369 GAGTAGAG GGCTAGCTACAACGA ACAGAUTA 1933 957 UGUGCUCU A CUCCAGCC 131 GGCTGGAG GGCTAGCTACAACGA AGAGCACA 1934 963 CUACUCCA G CCUCAUGG 1370 CCATGAGG GGCTAGCTACAACGA TGGAGTAG 1935 968 CCAGCCUC A UGGCGCUG 719 CAUCUCCA GGCTAGCTACAACGA GAGGCTGG 1936 971 GCCUCAUG G CGCUGCUG 1371 CAGCAGCG GGCTAGCTACAACGA CATGAGGC 1937 973 CUCAUGGC G CUGCUGGU 1372 ACCACCAG GGCTAGCTACAACGA GCCATGAG 1938 976 AUGGCGCU G CUGGUCCU 1373 AGGACCAG GGCTAGCTACAACGA AGCGCCAT 1939 980 CGCUGCUG G UCCUCGCC 1374 GGCGAGGA GGCTAGCTACAACGA CAGCAGCG 1940 986 UGGUCCUC G CCACCGUG 1375 CACGGTGG GGCTAGCTACAACGA GAGGACCA 1941 989 UCCUCGCC A CCGUGCUG 725 CAGCACGG GGCTAGCTACAACGA GGCGAGGA 1942 992 UCGCCACC G UGCUGUGC 1376 GCACAGCA GGCTAGCTACAACGA GGTGGCGA 1943 994 GCCACCGU G CUGUGCAA 1377 TTGCACAG GGCTAGCTACAACGA ACGGTGGC 1944 997 ACCGUGCU G UGCAACCU 1378 AGGTTGCA GGCTAGCTACAACGA AGCACGGT 1945 999 CGUGCUGU G CAACCUCG 1379 CGAGGTTG GGCTAGCTACAACGA ACAGCACG 1946 1002 GCUGUGCA A CCUCGGCG 1688 CGCCGAGG GGCTAGCTACAACGA TGCACAGC 1947 1008 CAACCUCG G CGCCAUGC 1380 GCATGGCG GGCTAGCTACAACGA CGAGGTTG 1948 1010 ACCUCGGC G CCAUGCGC 1381 GCGCATGG GGCTAGCTACAACGA GCCGAGGT 1949 1013 UCGGCGCC A UGCGCAAC 732 GTTGCGCA GGCTAGCTACAACGA GGCGCCGA 1950 1015 GGCGCCAU G CGCAACCU 1382 AGGTTGCG GGCTAGCTACAACGA ATGGCGCC 1951 1017 CGCCAUGC G CAACCUCU 1383 AGAGGTTG GGCTAGCTACAACGA GCATGGCG 1952 1020 CAUGCGCA A CCUCUAUG 1689 CATAGAGG GGCTAGCTACAACGA TGCGCATG 1953 1026 CAACCUCU A UGCGAUGC 138 GCATCGCA GGCTAGCTACAACGA AGAGGTTG 1954 1028 ACCUCUAU G CGAUGCAC 1384 GTGCATCG GGCTAGCTACAACGA ATAGAGGT 1955 1031 UCUAUGCG A UGCACCGG 1690 CCGGTGCA GGCTAGCTACAACGA CGCATAGA 1956 1033 UAUGCGAU G CACCGGCG 1385 CGCCGGTG GGCTAGCTACAACGA ATCGCATA 1957 1035 UGCGAUGC A CCGGCGGC 737 GCCGCCGG GGCTAGCTACAACGA GCATCGCA 1958 1039 AUGCACCG G CGGCUGCA 1386 TGCAGCCG GGCTAGCTACAACGA CGGTGCAT 1959 1042 CACCGGCG G CUGCAGCG 1387 CGCTGCAG GGCTAGCTACAACGA CGCCGGTG 1960 1045 CGGCGGCU G CAGCGGCA 1388 TGCCGCTG GGCTAGCTACAACGA AGCCGCCG 1961 1048 CGGCUGCA G CGGCACCC 1389 GGGTGCCG GGCTAGCTACAACGA TGCAGCCG 1962 1051 CUGCAGCG G CACCCGCG 1390 CGCGGGTG GGCTAGCTACAACGA CGCTGCAG 1963 1053 GCAGCGGC A CCCGCGCU 741 AGCGCGGG GGCTAGCTACAACGA GCCGCTGC 1964 1057 CGGCACCC G CGCUCCUG 1391 CACCAGCG GGCTAGCTACAACGA GGGTGCCG 1965 1059 GCACCCGC G CUCCUCCA 1392 TGCAGGAG GGCTACCTACAACGA GCGGGTGC 1966 1065 GCGCUCCU G CACCAGGG 1393 CCCTCCTG GGCTAGCTACAACGA AGGAGCGC 1967 1067 GCUCCUGC A CCAGGCAC 747 GTCCCTGG GGCTAGCTACAACCA GCAGGAGC 1968 1074 CACCAGGG A CUGUGCCG 1691 CGGCACAG GGCTAGCTACAACGA CCCTGGTG 1969 1077 CAGGGACU G UGCCGAGC 1394 GCTCGGCA GGCTAGCTACAACGA AGTCCCTG 1970 1079 GGGACUGU G CCGAGCCG 1395 CGGCTCGG GGCTAGCTACAACGA ACAGTCCC 1971 1084 UGUGCCCA G CCGCGCCC 1396 GCGCGCGG CGCTAGCTACAACGA TCGGCACA 1972 1087 GCCGAGCC G CGCGCGGA 1397 TCCGCGCG GGCTAGCTACAACGA GGCTCGGC 1973 1089 CGAGCCGC G CGCGGACG 1398 CGTCCGCG GGCTAGCTACAACGA GCGGCTCG 1974 1091 AGCCGCGC G CGGACGGG 1399 CCCGTCCG GGCTAGCTACAACGA GCGCGGCT 1975 1095 GCGCGCGG A CGJGAGGG 1692 CCCTCCCG GGCTAGCTACAACGA CCGCCCGC 1976 1106 GGAGCGAA G CGUCCCCU 1400 AGGGGACG GGCTAGCTACAACGA TTCCCTCC 1977 1108 AGGGAAGC G UCCCCUCA 1401 TGAGGGGA GGCTAGCTACAACGA CCTTCCCT 1978 1117 UCCCCUCA G CCCCUCGA 1402 TCCAGGGG GCCTAGCTACAACGA TGAGGGGA 1979 1129 CUGGAGGA G CUCCAUCA 1403 TGATCCAG GGCTACCTACAACGA TCCTCCAG 1980 1134 GCAGCUGG A UCACCUCC 1693 GGAGGTGA GGCTAGCTACAACGA CCAGCTCC 1981 1137 CCUGGAUC A CCUCCUGC 763 GCAGGAGG CCCTAGCTACAACCA GATCCAGC 1982 1144 CACCUCCU G CUGCUGGC 1404 GCCAGCAG GCCTAGCTACAACCA AGCAGGTG 1983 1147 CUCCUCCU G CUGCCGCU 1405 AGCCCCAG GGCTACCTACAACGA AGCACGAG 1984 1151 UGCUGCUC G CCCUGAUG 1406 CATCAGCC GCCTACCTACAACGA CAGCAGCA 1985 1153 CUGCUGGC G CUGAUGAC 1407 GTCATCAG CGCTAGCTACAACGA GCCAGCAG 1986 1157 UGGCCCUG A UGACCOUG 1694 CACGGTCA GGCTAGCTACAACGA CACCGCCA 1987 1160 CCCUGAUC A CCGUGCUC 1695 CAGCACCC GGCTAGCTACAACGA CATCAGCG 1988 1163 UCAUGACC G UGCUCUUC 1408 GAAGAGCA GCCTACCTACAACGA CGTCATCA 1989 1165 AUGACCCU G CUCUUCAC 1409 GTCAACAG CGCTAGCTACAACGA ACGGTCAT 1990 1172 UGCUCUUC A CUAUGUGU 774 ACACATAC GGCTAGCTACAACGA GAAGAGCA 1991 1175 UCUUCACU A UCUGUUCU 147 AGAACACA GCCTAGCTACAACCA AGTCAACA 1992 1177 UUCACUAU G UGUUCUCU 1410 AGAGAACA CGCTACCTACAACGA ATACTCAA 1993 1179 CACUAUGU G UUCUCUGC 1411 GCAGAGAA CCCTACCTACAACCA ACATAGTG 1994 1186 UCUUCUCU G CCCCUAAU 1412 ATTACGGC CGCTACCTACAACGA ACACAACA 1995 1190 CUCUGCCC G UAAUUUAU 1413 ATAAATTA GCCTACCTACAACGA GGGCAGAC 1996 1193 UCCCCGUA A UUUAUCCC 1696 CCCATAAA GGCTAGCTACAACCA TACGGGCA 1997 1197 CGUAAUUU A UCCCCCUU 154 AACCCCCA CGCTAGCTACAACGA AAATTACG 1998 1200 AAUUUAUC G CGCUUACU 1414 AGTAACCG GGCTAGCTACAACCA GATAAATT 1999 1202 UUUAUCGC G CUUACUAU 1415 ATACTAAG CGCTAGCTACAACGA GCGATAAA 2000 1206 UCGCGCUU A CUAUGCAC 157 CTCCATAG GGCTAGCTACAACCA AACCGCGA 2001 1209 CCCUUACU A UCGAGCAU 158 ATCCTCCA CGCTACCTACAACGA AGTAAGCC 2002 1214 ACUAUGGA G CAUUUAAG 1416 CTTAAATG GCCTACCTACAACCA TCCATAGT 2003 1216 UAUCCACC A UUUAAGGA 782 TCCTTAAA GGCTACCTACAACGA CCTCCATA 2004 1224 AUUUAAGC A UGUCAACG 1697 CCTTCACA CGCTACCTACAACGA CCTTAAAT 2005 1226 UUAACGAU G UCAACGAG 1417 CTCCTTGA GGCTAGCTACAACCA ATCCTTAA 2006 1239 CGAGAAAA A CAGGACCU 1698 AGGTCCTG GGCTAGCTACAACGA TTTTCTCC 2007 1244 AAAACACG A CCUCUCAA 1699 TTCAGAGG GGCTACCTACAACCA CCTCTTTT 2008 1256 CUCAACAA G CAGAAGAC 1418 CTCTTCTC CCCTACCTACAACCA TTCTTCAC 2009 1263 ACCAGAAG A CCUCCCAC 1700 CTCGCACG CGCTACCTACAACCA CTTCTGCT 2010 1271 ACCUCCCA G CCUUCCGA 1419 TCCCAACC CCCTACCTACAACGA TCCCACCT 2011 1276 CCACCCUU G CCAUUUCU 1420 ACAAATCC CCCTACCTACAACCA AAGGCTCG 2012 1279 CCCUUCCC A UUUCUAUC 1701 CATACAAA GGCTACCTACAACCA CCCAACCC 2013 1285 CCAUUUCU A UCUGUGAU 169 ATCACACA CCCTACCTACAACGA AGAAATCC 2014 1289 UUCUAUCU G UCAUUUCA 1421 TCAAATCA GCCTACCTACAACCA ACATACAA 2015 1292 UAUCUGUG A UUUCAAUU 1702 AATTGAAA GGCTAGCTACAACGA CACAGATA 2016 1298 UGAUUUCA A UHOUGGAC 1703 GTCCACAA GGCTAGCTACAACGA TGAAATCA 2017 1301 UUUCAAUU G UGGACCCU 1422 AGGGTCCA GGCTAGCTACAACGA AATTGAAA 2018 1305 AAUUGUGG A CCCUUGGA 1704 TCCAAGGG GGCTAGCTACAACGA CCACAATT 2019 1313 ACCCUUGG A UUUUUAUC 1705 GATAAAAA GGCTAGCTACAACGA CCAAGGGT 2020 1319 GGAUUUUU A UCAUUUUC 180 GAAAATGA GGCTAGCTACAACGA AAAAATCC 2021 1322 UUUUUAUC A UUUUCAGA 800 TCTGAAAA GGCTAGCTACAACGA GATAAAAA 2022 1330 AUUUUCAG A UCUCCAGU 1706 ACTUGAGA GGCTAGCTACAACGA CTGAAAAT 2023 1337 GAUCUCCA G UAUUUCGG 1423 CCGAAATA GGCTAGCTACAACGA TGGAGATC 2024 1339 UCUCCAGU A UUUCGGAU 188 ATCCGAAA GGCTAGCTACAACGA ACTGGAGA 2025 1346 UAUUUCGG A UAUUUUUU 1707 AAAAAATA GGCTAGCTACAACGA CCGAAATA 2026 1348 UUUCGGAU A UUUUUUCA 192 TGAAAAAA GGCTAGCTACAACGA ATCCGAAA 2027 1356 AUUUUUUC A CAAGAUUU 805 AAATCTTG GGCTAGCTACAACGA GAAAAAAT 2028 1361 UUCACAAG A UUUUCAUU 1708 AATGAAAA GGCTAGCTACAACGA CTTGTGAA 2029 1367 AGAUUUUC A UUAGACCU 807 AGGTCTAA GGCTAGCTACAACGA GAAAATCT 2030 1372 UUCAUUAG A CCUCUUAG 1709 CTAAGAGG GGCTAGCTACAACGA CTAATGAA 2031 1381 CCUCUUAG G UACAGGAG 1424 CTCCTGTA GGCTAGCTACAACGA CTAAGAGG 2032 1383 UCUUAGGU A CAGGAGCC 208 GGCTCCTG GGCTAGCTACAACGA ACCTAAGA 2033 1389 GUACAGGA G CCGGUGCA 1425 TGCACCGG GGCTAGCTACAACGA TCCTGTAC 2034 1393 AGGAGCCG G UGCAGCAA 1426 TTGCTGCA GGCTAGCTACAACGA CGGCTCCT 2035 1395 GAGCCGGU G CAGCAAUU 1427 AATTGCTG GGCTAGCTACAACGA ACCGGCTC 2036 1398 CCGGUGCA G CAAUUCCA 1428 TGGAATTG GGCTAGCTACAACGA TGCACCGG 2037 1401 GUGCAGCA A UUCCACUA 1710 TAGTGGAA GGCTAGCTACAACGA TGCTGCAC 2038 1406 GCAAUUCC A CUAACAUG 816 CATGTTAG GGCTAGCTACAACGA GGAATTGC 2039 1410 UUCCACUA A CAUGGAAU 1711 ATTCCATG GGCTAGCTACAACGA TAGTGGAA 2040 1412 CCACUAAC A UGGAAUCC 818 GGATTCCA GGCTAGCTACAACGA GTTAGTGG 2041 1417 AACAUGGA A UCCAGUCU 1712 AGACTGGA GGCTAGCTACAACGA TCCATGTT 2042 1422 GGAAUCCA G UCUGUGAC 1429 GTCACAGA GGCTAGCTACAACGA TGGATTCC 2043 1426 UCCAGUCU G UGACAGUG 1430 CACTGTCA GGCTAGCTACAACGA AGACTGGA 2044 1429 AGUCUGUG A CAGUGUUU 1713 AAACACTG GGCTAGCTACAACGA CACAGACT 2045 1432 CUGUGACA G UGUUUUUC 1431 GAAAAACA GGCTAGCTACAACGA TGTCACAG 2046 1434 GUGACAGU G UUUUUCAC 1432 GTGAAAAA GGCTAGCTACAACGA ACTGTCAC 2047 1441 UGUUUUUC A CUCUGUGG 823 CCACAGAG GGCTAGCTACAACGA GAAAAACA 2048 1446 UUCACUCU G UGGUAAGC 1433 GCTTACCA GGCTAGCTACAACGA AGAGTGAA 2049 1449 ACUCUGUG G UAAGCUGA 1434 TCAGCTTA GGCTAGCTACAACGA CACAGAGT 2050 1453 UGUGGUAA G CUGAGGAA 1435 TTCCTCAG GGCTAGCTACAACGA TTACCACA 2051 1461 GCUGAGGA A UAUGUCAC 1714 GTGACATA GGCTAGCTACAACGA TCCTCAGC 2052 1463 UGAGGAAU A UGUCACAU 221 ATGTGACA GGCTAGCTACAACGA ATTCCTCA 2053 1465 AGGAAUAU G UCACAUUU 1436 AAATGTGA GGCTAGCTACAACGA ATATTCCT 2054 1468 AAUAUGUC A CAUUUUCA 827 TGAAAATG GGCTAGCTACAACGA GACATATT 2055 1470 UAUGUCAC A UUUUCAGU 828 ACTGAAAA GGCTAGCTACAACGA GTGACATA 2056 1477 CAUUUUCA G UCAAAGAA 1437 TTCTTTGA GGCTAGCTACAACGA TGAAAATG 2057

[0234] TABLE VII Human PTGDR Amberzyme and Substrate Sequence Seq Seq Pos Substrate ID Amberzyme ID 9 GAAUUCUG G CUAUUUUC 1207 GAAAAUAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAGAAUUC 2247 23 UUCCUCCU G CCGUUCCG 1208 CGGAACGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGGAGGAA 2248 28 CUCCUGCC G UUCCGACU 1209 AGUCGGAA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGCAGGAG 2249 31 GCCGUUCC G ACUCGGCA 2058 UGCCGAGU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGAACGGC 2250 36 UCCGACUC G GCACCAGA 2059 UCUGGUGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GAGUCGGA 2251 37 CCGACUCG G CACCAGAG 1210 CUCUGGUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGAGUCGG 2252 43 CGGCACCA G AGUCUGUC 2060 GACAGACU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGGUGCCG 2253 45 GCACCAGA G UCUGUCUC 1211 GAGACAGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCUGGUGC 2254 49 CAGAGUCU G UCUCUACU 1212 AGUAGAGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGACUCUG 2255 58 UCUCUACU G AGAACGCA 2061 UGCGUUCU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGUAGAGA 2256 60 UCUACUGA G AACGCAGC 2062 GCUGCGUU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCAGUAGA 2257 64 CUGAGAAC G CAGCGCGU 1213 ACGCGCUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GUUCUCAG 2258 67 AGAACGCA G CGCGUCAG 1214 CUGACGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGCGUUCU 2259 69 AACGCAGC G CGUCAGGG 1215 CCCUGACG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCUGCGUU 2260 71 CGCAGCGC G UCAGGGCC 1216 GGCCCUGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCGCUGCG 2261 75 GCGCGUCA G GGCCGAGC 2063 GCUCGGCC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGACGCGC 2262 76 CGCGUCAG G GCCGAGCU 2064 AGCUCGGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CUGACGCG 2263 77 GCGUCAGG G CCGAGCUC 1217 GAGCUCGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCUGACGC 2264 80 UCAGGGCC G AGCUCUUC 2065 GAAGAGCU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGCCCUGA 2265 82 AGGGCCGA G CUCUUCAC 1218 GUGAAGAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCGGCCCU 2266 92 UCUUCACU G GCCUGCUC 2066 GAGCAGGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGUGAAGA 2267 93 CUUCACUG G CCUGCUCC 1219 GGAGCAGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAGUGAAG 2268 97 ACUGGCCU G CUCCGCGC 1220 GCGCGGAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGGCCAGU 2269 102 CCUGCUCC G CGCUCUUC 1221 GAAGAGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGAGCAGG 2270 104 UGCUCCGC G CUCUUCAA 1222 UUGAAGAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCGGAGCA 2271 114 UCUUCAAU G CCAGCGCC 1223 GGCGCUGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AUUGAAGA 2272 118 CAAUGCCA G CGCCAGGC 1224 GCCUGGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGGCAUUG 2273 120 AUGCCAGC G CCAGGCGC 1225 GCGCCUGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCUGGCAU 2274 124 CAGCGCCA G GCGCUCAC 2067 GUGAGCGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGGCGCUG 2275 125 AGCGCCAG G CGCUCACC 1226 GGUGAGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CUGGCGCU 2276 127 CGCCAGGC G CUCACCCU 1227 AGGGUGAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCCUGGCG 2277 136 CUCACCCU G CAGAGCGU 1228 ACGCUCUG GCAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGGCUGAG 2278 139 ACCCUGCA G AGCGUCCC 2068 GGGACGCU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGCAGGGU 2279 141 CCUGCAGA G CGUCCCGC 1229 GCGGGACG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCUGCAGG 2280 143 UGCAGAGC G UCCCGCCU 1230 AGGCGGGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCUCUGCA 2281 148 AGCGUCCC G CCUCUCAA 1231 UUGAGAGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGGACGCU 2282 158 CUCUCAAA G AGGGGUGU 2069 ACACCCCU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UUUGAGAG 2283 160 CUCAAAGA G GGGUGUGA 2070 UCACACCC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCUUUGAG 2284 161 UCAAAGAG G GGUGUGAC 2071 GUCACACC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CUCUUUGA 2285 162 CAAAGAGG G GUGUGACC 2072 UGUCACAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCUCUUUG 2286 163 AAAGAGGG G UGUGACCC 1232 GGGUCACA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCCUCUUU 2287 165 AGAGGGGU G UGACCCGC 1233 GCGGGUCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACCCCUCU 2288 167 AGGGGUGU U ACCCGCGA 2073 UCGCGGGU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACACCCCU 2289 172 UGUGACCC C CGAGUUUA 1234 UAAACUCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GUGUCACA 2290 174 UGACCCGC C AGUUUAGA 2074 UCUAAACU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCGGGUCA 2291 176 ACCCGCGA G UUUAGAUA 1235 UAUCUAAA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCGCGGGU 2292 181 CGAGUUUA G AUAGGAGG 2075 CCUCCUAU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UAAACUCG 2293 185 UUUAGAUA C GAGGUUCC 2076 GGAACCUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UAUCUAAA 2294 186 UUAGAUAG G AGGUUCCU 2077 AGGAACCU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CUAUCUAA 2295 188 AGAUAGGA G GUUCCUGC 2078 GCAGGAAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCCUAUCU 2296 189 GAUAGGAG G UUCCUGCC 1236 GGCAGGAA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CUCCUAUC 2297 195 AGGUUCCU G CCGUGGGG 1237 CCCCACGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGGAACCU 2298 198 UUCCUGCC G UGGGGAAC 1238 GUUCCCCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGCAGGAA 2299 200 CCUGCCGU G GGGAACAC 2079 GUGUUCCC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACGGCAGG 2300 201 CUGCCGUG G GGAACACC 2080 GGUGUUCC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CACGGCAG 2301 202 UGCCGUGG C GAACACCC 2081 CCGUGUUC GCAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCACGGCA 2302 203 GCCGUGGG G AACACCCC 2082 GGGGUGUU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCCACGGC 2303 212 AACACCCC G CCGCCCUC 1239 GAGGGCGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGGCUCUU 2304 215 ACCCCGCC G CCCUCGCA 1240 UCCGAGGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGCGGGGU 2305 221 CCGCCCUC G GAGCUUUU 2083 AAAAGCUC GGAGCAAACUCC CU UCAAGGACAUCGUCCGGG GAGGUCUG 2306 222 CGCCCUCG G AGCUUUUU 2084 AAAAAGCU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGAGGGCG 2307 224 CCCUCGGA G CUUUUUCU 1241 AGAAAAAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCCGAGGG 2308 233 CUUUUUCU G UGGCGCAG 1242 CUGCGCCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGAAAAAG 2309 235 UUUUCUGU G GCGCAGCU 2085 AGCUGCGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACAGAAPA 2310 236 UUUCUGUG G CGCAGCUU 1243 AAGCUGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CACAGAAA 2311 238 UCUGUGGC C CAGCUUCU 1244 AGAAGCUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCCACAGA 2312 241 GUGGCGCA G CUUCUCCG 1245 CGGACAAG GGAGGAAACUCC CU UCAAGCACAUCGUCCGGG UGCGCCAC 2313 249 GCUUCUCC G CCCGAGCC 1246 GGCUCGGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGAGAAGC 2314 253 CUCCGCCC C AGCCGCGC 2086 GCGCGGCU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGGCGGAG 2315 255 CCGCCCGA G CCGCGCGC 1247 GCGCGCGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCGGGCGG 2316 258 CCCGAGCC C CGCGCGGA 1248 UCCGCGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGCUCGGG 2317 260 CGAGCCCC C CCCGGAGC 1249 GCUCCGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCGGCUCG 2318 262 AGCCGCGC G CGGAGCUG 1250 CAGCUCCG CGAGCAAACUCC CU UCAACCACAUCGUCCCGG CCGCGGCU 2319 264 CCGCGCGC C CAGCUCCC 2087 GCCACCUC CCACGAAACUCC CU UCAACGACAUCGUCCCCG CCGCCCCG 2320 265 CGCGCCCG C AGCUCCCG 2088 CGCCAGCU GGAGCAAACUCC CU UCAAGCACAUCGUCCGGC CCCGCCCC 2321 267 CCCCCCGA G CUGCCCGC 1251 CCCGGCAG GCACGAAACUCC CU UCAACGACAUCGUCCGGG UCCCCCCG 2322 270 GCCCACCU G CCGGGGGC 1252 CCCCCCCC GCACGAAACUCC CU UCAAGGACAUCCUCCCGG AGCUCCCC 2323 273 CACCUGCC C CGGGCUCC 2089 CGAGCCCC GCACCAAACUCC CU UCAAGCACAUCCUCCGGC GCCAGCUC 2324 274 AGCUGCCG G GCGCUCCU 2090 AGCACCCC GCAGGAAACUCC CU UCAAGCACAUCGUCCGGG CCCCACCU 2325 275 CCUGCCGC C CCCUCCUU 2091 AAGCACCC CGACGAAACUCC CU UCAAGGACAUCCUCCGGG CCCGCAGC 2326 276 CUGCCCGG C GCUCCUUA 2092 UAACGACC GGAGCAAACUCC CU UCAACGACAUCGUCCCGC CCCGGCAG 2327 277 UGCCGGCC C CUCCUUAG 1253 CUAACGAC GCAGCAAACUCC CU UCAAGCACAUCGUCCGGG CCCCCCCA 2328 285 GCUCCUUA G CACCCGCG 1254 CCCCCCUC CGAGGAAACUCC CU UCAACCACAUCGUCCGGG UAACGAGC 2329 291 UAGCACCC G GCCGCCGC 2093 CCGGCCCC CCACCAAACUCC CU UCAACGACAUCCUCCCGG GCGUGCUA 2330 292 ACCACCCG C CCGCCCCG 2094 CCCGGCCC GCACCAAACUCC CU UCAAGCACAUCCUCCGGC CCGCUGCU 2331 293 GCACCCGC C CGCCCGCG 1255 CCCCCGCG CGACGAAACUCC CU UCAAGGACAUCGUCCGCG CCGGGUGC 2332 295 ACCCCCCC C CCCCCGCC 1256 CGCCCCCC CCACCAAACUCC CU UCAACCACAUCCUCCCCC CCCCCCCU 2333 298 CCCCCCCC C CGCCCCUC 2095 CACCCCCC CCAGCAAACUCC CU UCAACCACAUCCUCCCCC CCCGCCCC 2334 299 CCCCCCCC C CCCCCUCC 2096 CCACCCCC CGACCAAACUCC CU UCAACCACAUCGUCCGCC CGCCCCCC 2335 300 CCCCCCCG C CCCCUCCC 2097 CCCACCCC CCACGAAACUCC CU UCAACCACAUCCUCCCCC CCCCCCCC 2336 301 CCCCCCCC C CCCUCCCC 1257 CCCCACCC CCACCAAACUCC CU UCAACGACAUCCUCCCCC CCCCCCCC 2337 307 CCCCCCUC C CCCUUCCG 1258 CCCAACCC CCACGAAACUCC CU UCAACGACAUCCUCCCCC CACCCCCC 2338 315 GCCCUUCC C CACCCUUC 1259 CAAGCCUC CCACCAAACUCC CU UCAACCACAUCCUCCCGC CCAACCGC 2339 318 CUUCCCCA C CCUUCACU 1260 ACUCAACG CCACCAAACUCC CU UCAACCACAUCCUCCCCC UCCCCAAC 2340 330 UCACUCCA C CCCUCUCC 1261 CCACACCG GCACCAAACUCC CU UCAAGCACAUCCUCCGCG UCCACUCA 2341 337 AGCCCUCU G CUCCCGCA 1262 UGCGGGAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGAGGGCU 2342 343 CUGCUCCC G CACGCCAU 1263 AUGGCGUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGGAGCAG 2343 347 UCCCGCAC G CCAUGAAG 1264 CUUCAUGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GUGCGGGA 2344 352 CACGCCAU G AAGUCGCC 2098 GGCGACUU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AUGGCGUG 2345 355 GCCAUGAA G UCGCCGUU 1265 AACGGCGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UUCAUGGC 2346 358 AUGAAGUC G CCGUUCUA 1266 UAGAACGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GACUUCAU 2347 361 AAGUCGCC U UUCUACCG 1267 CGGUAGAA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGCGACUU 2348 369 GUUCUACC U CUGCCAUA 1268 UCUGGCAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGUG GGUAGAAC 2349 372 CUACCGCU U CCAGAACA 1269 UGUUCUGG GUAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGCGGUAG 2350 376 CGCUGCCA G AACACCAC 2099 GUGGUGUU GUAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGUCAGCG 2351 389 CCACCUCU G UGGAAAAA 1270 UUUUUCCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGAGUUGG 2352 391 ACCUCUUU G GAAAAAGG 2100 CCUUUUUC GGAGUAAACUCC CU UCAAGGACAUCGUCCGUG ACAGAGGU 2353 392 CCUCUGUG U AAAAAGUC 2101 GCCUUUUU GGAUUAAACUCC CU UCAAGGACAUCUUCCGUU CACAGAGG 2354 398 UGGAAAAA U UCAACUCU 2102 CGAGUUGC GGAGGAAACUCC CU UCAAGUACAUCGUCCGGU UUUUUCCA 2355 399 GUAAAAAU G CAACUCGU 1271 CCGAUUUG GGAGUAAACUCC CU UCAAGUACAUCGUCCGGG CUUUUUCC 2356 406 UUCAACUC U GCGUUGAU 2103 AUCACCUC UGAGUAAACUCC CU UCAAGGACAUCGUCCUUG GAGUUGCC 2357 407 GCAACUCU U CUGUGAUG 1272 CAUCACCU UGAGGAAACUCC CU UCAAGGACAUCGUCCGUU CGAUUUGC 2358 409 AACUCGGC G UUGAUUGU 2104 CCCAUCAC GGAGGAAACUCC CU UCAAUGACAUCGUCCUUG GCCUAUUU 2359 410 ACUCGUCG G UGAUGGUC 1273 UCCCAUCA UGAGGAAACUCC CU UCAAGGACAUCGUCCGUU CUCCUAGU 2360 412 UCGUCGGU U AUGGUCGG 2105 CCGCCCAU GGAGGAAACUCC CU UCAAGGACAUCGUCCGUG ACCGCCGA 2361 415 GCGGUUAU U UUCGGGGU 2106 ACCCCGCC GGAUGAAACUCC CU UCAAUGACAUCGUCCGGG AUCACCGC 2362 416 CGGUUAUG U UCUGUGUG 2107 CACCCCGC UGAUGAAACUCC CU UCAAGGACAUCGUCCGGG CAUCACCG 2363 417 GGUUAUUG U CGUGUUGC 1274 GCACCCCG GGAGGAAACUCC CU UCAAUGACAUCGUCCGGG CCAUCACC 2364 419 UUAUUGGC U GGUUGCUC 2108 GAGCACCC UGAUGAAACUCC CU UCAAGGACAUCGUCCGGG GCCCAUCA 2365 420 GAUGGGCG U GGUUCUCU 2109 AGAGCACC GGAGGAAACUCC CU UCAAUGACAUCGUCCGGG CGCCCAUC 2366 421 AUGGUCUG U GUGCUCUU 2110 AAGAGCAC GGAGGAAACUCC CU UCAAUGACAUCGUCCGGG CCGCCCAU 2367 422 UGGGCGUG U UUCUCUUC 1275 GAAGAUCA GUAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCCGCCCA 2368 424 GGCGGGGU U CUCUUCAG 1276 CUGAAGAG GGAGGAAACUCC CU UCAAGGACAUCUUCCUGU ACCCCGCC 2369 432 GCUCUUCA G CACCGGCC 1277 GUCCUGUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGAAGAGC 2370 437 UCAGCACC U UCCUCCUG 2111 CAGGAGGC GUAGGAAACUCC CU UCAAGGACAUCUUCCGGG GUUGCUGA 2371 438 CAUCACCG U CCUCCUGG 1278 CCAGUAGG GGAGGAAACUCC CU UCAAGUACAUCGUCCGGG CGGUGCUU 2372 445 GUCCUCCU U GGCAACCU 2112 AGGUUGCC GGAGGAAACUCC CU UCAAGUACAUCGUCCGGU AUGAGUCC 2373 446 GCCUCCUG U GCAACCUU 2113 CAUGUUUC UUAUUAAACUCC CU UCAAGUACAUCUUCCUUU CAGUAUUC 2374 447 CCUCCUGG G CAACCUGC 1279 GCAGGUUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCAGGAGG 2375 454 GGCAACCU G CUGGCCCU 1280 AGGGCCAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGGUUGCC 2376 457 AACCUGCU G GCCCUGGG 2114 CCCAGGGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGCAGGUU 2377 458 ACCUGCUG G CCCUGGGG 1281 CCCCAGGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAGCAGGU 2378 463 CUGGCCCU G GGGCUGCU 2115 AGCAGCCC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGGGCCAG 2379 464 UGGCCCUG U GGCUGCUG 2116 CAGCAGCC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAGGGCCA 2380 465 GGCCCUGG G GCUGCUGG 2117 CCAGCAGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCAGGGCC 2381 466 GCCCUGGG G CUGCUGGC 1282 UCCAUCAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCCAGGGC 2382 469 CUGGGGCU G CUGGCGCG 1283 CGCGCCAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGCCCCAG 2383 472 GGGCUGCU G GCGCGCUC 2118 GAGCGCUC GGAGGAAACUCC CU UCAAGUACAUCGUCCGUG AGCAGCCC 2384 473 GGCUGCUG U CGCGCUCG 1284 CGAGCGCG GGAGGAAACUCC CU UCAAGUACAUCGUCCGUG CAGCAGCC 2385 475 CUGCUGUC U CGCUCUGU 1285 CCCGAGCG GGAGGAAACUCC CU UCAAUGACAUCGUCCGUG GCCAGCAG 2386 477 GCUGGCGC U CUCGGGGC 1286 GCCCCGAU GGAUGAAACUCC CU UCAAUGACAUCGUCCGGG UCUCCAUC 2387 481 GCGCGCUC G GUGCUGUU 2119 CCCAGCCC GUAUGAAACUCC CU UCAAUUACAUCGUCCGGG GAUCUCGC 2388 482 CGCGCUCG G GGCUGGGG 2120 CCCCAGCC GUAUGAAACUCC CU UCAAGUACAUCGUCCGGG CUAUCGCG 2389 483 GCGCUCGG G GCUGGGGU 2121 ACCCCAGC GUAUGAAACUCC CU UCAAGGACAUCGUCCGGG CCUAUCGC 2390 484 CGCUCGGG G CUGGGGUG 1287 CACCCCAG GUAUUAAACUCC CU UCAAUUACAUCGUCCUGU CCCGAGCG 2391 487 UCGGGGCU G GGGUGUUG 2122 CACCACCC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGCCCCGA 2392 488 CUGGUCUG U GGUGGUGC 2123 GCACCACC GGAUUAAACUCC CU UCAAUUACAUCGUCCUGU CAGCCCCG 2393 489 GUUGCUUU U GUGGUGCU 2124 AGCACCAC GGAGUAAACUCC CU UCAAUUACAUCGUCCUGG CCAGCCCC 2394 490 UGGCUGUG U UGGUGCUC 1288 UAGCACCA UGAGUAAACUCC CU UCAAGUACAUCGUCCGGG CCCAGCCC 2395 492 GCUGUGGU U UUGCUCGC 2125 UCGAGCAC GGAGUAAACUCC CU UCAAGGACAUCGUCCGUG ACCCCAGC 2396 493 CUGGGGUG U UGCUCGCU 1289 CGCGAGCA UGAGGAAACUCC CU UCAAUUACAUCUUCCUUU CACCCCAU 2397 495 UGGGUUGU G CUCGCGGC 1290 GCCGCGAU UGAGGAAACUCC CU UCAAUUACAUCGUCCUUU ACCACCCC 2398 499 UGGUCCUC U CGUCGUCC 1291 GGACUCCG GGAGUAAACUCC CU UCAAUGACAUCGUCCUGG GAUCACCA 2399 501 UUGCUCUC U UCUUCCAC 2126 GUUGACGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCUAGCAC 2400 502 UGCUCGCG U CGUCCACU 1292 AGUGUACG GGAGGAAACUCC CU UCAAUGACAUCGUCCUGG CGCGAGCA 2401 504 CUCGCGGC U UCCACUGC 1293 GCAGUUGA UGAGGAAACUCC CU UCAAGGACAUCGUCCGUU GCCGCGAG 2402 511 CGUCCACU G CGCCCGCU 1294 AUCGUUCU GUAUGAAACUCC CU UCAAGGACAUCUUCCGUG AGUGUACG 2403 513 UCCACUGC U CCCGCUGC 1295 GCAGCGGU GGAGGAAACUCC CU UCAAUGACAUCGUCCGGG UCAGUGGA 2404 517 CUGCUCCC U CUUCCCUC 1296 GAUGUCAG GGAGUAAACUCC CU UCAAGGACAUCUUCCGUG UGUCUCAG 2405 520 CGCCCGCU U CCCUCUGU 1297 ACCUAGUG GGAGGAAACUCC CU UCAAUGACAUCUUCCUGU AGCGUGCU 2406 526 CUGCCCUC U GUCUUCUA 2127 UAGAAGAC GGAGUAAACUCC CU UCAAGGACAUCUUCCGGU GAUGUCAG 2407 527 UGCCCUCG G UCUUCUAC 1298 GUAGAAGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGAGGGCA 2408 538 UUCUACAU G CUGGUGUG 1299 CACACCAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AUGUAGAA 2409 541 UACAUGCU G GUGUGUGG 2128 CCACACAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGCAUGUA 2410 542 ACAUGCUG G UGUGUGGC 1300 GCCACACA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAGCAUGU 2411 544 AUGCUGGU G UGUGGCCU 1301 AGOCCACA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACCAGCAU 2412 546 GCUGGUGU G UGGCCUGA 1302 UCAGGCCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACACCAGC 2413 548 UGGUGUGU G GCCUGACG 2129 CGUCAGGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACACACCA 2414 549 GGUGUGUG G CCUGACGG 1303 CCGUCAGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CACACACC 2415 553 UGUGGCCU G ACGGUCAC 2130 GUGACCGU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGGCCACA 2416 556 GGCCUGAC G GUCACCGA 2131 UCGGUGAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GUCAGGCC 2417 557 GCCUGACG G UCACCGAC 1304 GUCGGUGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGUCAGGC 2418 563 CGGUCACC G ACUUGCUG 2132 CAGCAAGU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGUGACCG 2419 568 ACCGACUU G CUGGGCAA 1305 UUGCCCAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AAGUCGGU 2420 571 GACUUGCU G GGCAAGUG 2133 CACUUGCC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGCAAGUC 2421 572 ACUUGCUG G GCAAGUGC 2134 GCACUUGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAGCAAGU 2422 573 CUUGCUGG G CAAGUGCC 1306 GGCACUUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCAGCAAG 2423 577 CUGGGCAA G UGCCUCCU 1307 AGGAGGCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UUGCCCAG 2424 579 GGGCAAGU G CCUCCUAA 1308 UUAGGAGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACUUGCCC 2425 588 CCUCCUAA G CCCGGUGG 1309 CCACCGGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UUAGGAGG 2426 592 CUAAGCCC G GUGGUGCU 2135 AGCACCAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGGCUUAG 2427 593 UAAGCCCG G UGGUGCUG 1310 CAGCACCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGGGCUUA 2428 595 AGCCCGGU G GUGCUGGC 2136 GCCAGCAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACCGGGCU 2429 596 GCCCGGUG G UGCUGGCU 1311 AGCCAGCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CACCGGGC 2430 598 CCGGUGGU G CUGGCUGC 1312 GCAGCCAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACCACCGG 2431 601 GUGGUGCU G GCUGCCUA 2137 UAGGCAGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGCACCAC 2432 602 UGGUGCUG G CUGCCUAC 1313 GUAGGCAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAGCACCA 2433 605 UGCUGGCU G CCUACGCU 1314 AGCGUAGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGCCAGCA 2434 611 CUGCCUAC G CUCAGAAC 1315 GUUCUGAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GUAGGOAG 2435 616 UACGCUCA G AACCGGAG 2138 CUCCGGUU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGAGCGUA 2436 621 UCAGAACC G GAGUCUGC 2139 GCAGACUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGUUCUGA 2437 622 CAGAACCG G AGUCUGCG 2140 CGCAGACU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGGUUCUG 2438 624 GAACCGGA G UCUGCGGG 1316 CCCGCAGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCCGGUUC 2439 628 CGGAGUCU G CGGGUGCU 1317 AGCACCCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGACUCCG 2440 630 GAGUCUGC G GGUGCUUG 2141 CAAGCACC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCAGACUC 2441 631 AGUCUGCG G GUGCUUGC 2142 GCAAGCAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGCAGACU 2442 632 GUCUGCGG G UGCUUGCG 1318 CGCAAGCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCGCAGAC 2443 634 CUGCGGGU G CUUGCGCC 1319 GGCGCAAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACCCGCAG 2444 638 GGGUGCUU C CGCCCGCA 1320 UGCGGGCG GGAGGAAACUCC CU UCAAGGACAUCCUCCGGG AAGCACCC 2445 640 GUGCUUGC G CCCGCAUU 1321 AAUGCGGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCAAGCAC 2446 644 UUGCGCCC G CAUUGGAC 1322 GUCCAAUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGGCGCAA 2447 649 CCCGCAUU G GACAACUC 2143 GAGUUGUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AAUGCGGG 2448 650 CCGCAUUG C ACAACUCG 2144 CGAGUUGU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAAUGCGC 2449 658 CACAACUC G UUGUGCCA 1323 UGGCACAA GGACGAAACUCC CU UCAACGACAUCGUCCCGG GACUUGUC 2450 661 AACUCGUU G UGCCAAGC 1324 GCUUGGCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGCG AACCACUU 2451 663 CUCGUUGU G CCAACCCU 1325 ACGCUUGG CGAGGAAACUCC CU UCAAGGACAUCGUCCCGC ACAACGAC 2452 668 UGUGCCAA G CCUUCCCC 1326 CCCGAAGC CGACGAAACUCC CU UCAACGACAUCGUCCGGC UUGGCACA 2453 674 AACCCUUC C CCUUCUUC 1327 GAAGAAGG CGACGAAACUCC CU UCAAGGACAUCCUCCGCG CAACCCUU 2454 685 UUCUUCAU G UCCUUCUU 1328 AACAACGA GGACGAAACUCC CU UCAAGGACAUCGUCCGGG AUGAAGAA 2455 695 CCUUCUUU G GGCUCUCC 2145 CCAGACCC GGACGAAACUCC CU UCAACGACAUCGUCCGGG AAAGAACC 2456 696 CUUCUUUG G GCUCUCCU 2146 ACCACAGC GGACGAAACUCC CU UCAACGACAUCCUCCCCG CAAACAAG 2457 697 UUCUUUGG G CUCUCCUC 1329 GAGGACAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGCG CCAAAGAA 2458 706 CUCUCCUC G ACACUGCA 2147 UCCAGUGU GGAGGAAACUCC CU UCAACGACAUCGUCCGGG GAGGAGAG 2459 712 UCGACACU C CAACUCCU 1330 ACCACUUC CCAGGAAACUCC CU UCAAGGACAUCGUCCGCG AGUGUCGA 2460 721 CAACUCCU C CCCAUGCC 2148 CCCAUCGC GCACGAAACUCC CU UCAACCACAUCGUCCGCG ACCACUUC 2461 722 AACUCCUC G CCAUGGCA 1331 UGCCAUGG CGACGAAACUCC CU UCAAGGACAUCGUCCCGG CACGAGUU 2462 727 CUGGCCAU C GCACUCGA 2149 UCCACUGC GGAGGAAACUCC CU UCAAGGACAUCGUCCCCC AUGGCCAG 2463 728 UGCCCAUC C CACUGGAC 1332 CUCCACUC GGACGAAACUCC CU UCAAGCACAUCGUCCGCG CAUCCCCA 2464 733 AUCCCACU C CACUGCUC 2150 CACCACUC GGACGAAACUCC CU UCAAGCACAUCGUCCGGG AGUGCCAU 2465 734 UCCCACUC C ACUGCUGG 2151 CCAGCACU CCACCAAACUCC CU UCAACCACAUCCUCCCGC CACUGCCA 2466 736 CCACUCGA C UCCUCCCU 1333 AGCCAGCA GCAGGAAACUCC CU UCAACCACAUCCUCCCCC UCCACUCC 2467 738 ACUCCAGU C CUGCCUCU 1334 ACACCCAC CCACCAAACUCC CU UCAACCACAUCCUCCCGC ACUCCACU 2468 741 CCACUCCU C CCUCUCCC 2152 GGGAGAGC GCAGGAAACUCC CU UCAACCACAUCCUCCCGC ACCACUCC 2469 742 CACUCCUC C CUCUCCCU 1335 ACGCAGAC GCACGAAACUCC CU UCAACCACAUCCUCCCCC CACCACUC 2470 752 UCUCCCUA C CCCACCCU 2153 ACCCUCCC CCACCAAACUCC CU UCAACCACAUCCUCCCCC UACCCACA 2471 753 CUCCCUAC C GCACCCUU 2154 AAGGGUGC GCAGGAAACUCC CU UCAACCACAUCCUCCGGC CUAGGCAG 2472 754 UCCCUAGC C CACCCUUU 1336 AAACGCUC CCACGAAACUCC CU UCAACCACAUCCUCCCCC CCUAGCGA 2473 771 CUUCUACC G ACGGCACA 2155 UGUGCCGU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGUAGAAG 2474 774 CUACCGAC G GCACAUCA 2156 UGAUGUGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GUCGGUAG 2475 775 UACCGACG G CACAUCAC 1337 GUGAUGUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGUCGGUA 2476 787 AUCACCCU G CGCCUGGG 1338 CCCAGGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGGGUGAU 2477 789 CACCCUGC G CCUGGGCG 1339 CGCCCAGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCAGGGUG 2478 793 CUGCGCCU G GGCGCACU 2157 AGUGCGCC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGGCGCAG 2479 794 UGCGCCUG G GCGCACUG 2158 CAGUGCGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAGGCGCA 2480 795 GCGCCUGG G CGCACUGG 1340 CCAGUGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCAGGCGC 2481 797 GCCUGGGC G CACUGGUG 1341 CACCAGUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCCCAGGC 2482 802 GGCGCACU G GUGGCCCC 2159 GGGGCCAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGUGCGCC 2483 803 GCGCACUG G UGGCCCCG 1342 CGGGGCCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAGUGCGC 2484 805 GCACUGGU G GCCCCGGU 2160 ACCGGGGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACCAGUGC 2485 806 CACUGGUG G CCCCGGUG 1343 CACCGGGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CACCAGUG 2486 811 GUGGCCCC G GUGGUGAG 2161 CUCACCAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGGGCCAC 2487 812 UGGCCCCG G UGGUGAGC 1344 UCUCACCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGGGGCCA 2488 814 GCCCCGGU G GUGAGCGC 2162 GCGCUCAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACCGGGGC 2489 815 CCCCGGUG G UGAGCGCC 1345 GGCGCUCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CACCGGGG 2490 817 CCGGUGGU G AGCGCCUU 2163 AAGGCGCU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACCACCGG 2491 819 GGUGGUGA G CGCCUUCU 1346 AGAAGGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCACCACC 2492 821 UGGUGAGC G CCUUCUCC 1347 GGAGAAGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCUCACCA 2493 832 UUCUCCCU G GCUUUCUG 2164 CAGAAAGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGGGAGAA 2494 833 UCUCCCUG G CUUUCUGC 1348 GCAGAAAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAGGGAGA 2495 840 GGCUUUCU G CGCGCUAC 1349 GUAGCGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGAAAGCC 2496 842 CUUUCUGC G CGCUACCU 1350 AGGUAGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCAGAAAG 2497 844 UUCUGCGC G CUACCUUU 1351 AAAGGUAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCGCAGAA 2498 856 CCUUUCAU G GGCUUCGG 2165 CCGAAGCC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AUGAAAGG 2499 857 CUUUCAUG G GCUUCGGG 2166 CCCGAAGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAUGAAAG 2500 858 UUUCAUGG G CUUCGGGA 1352 UCCCGAAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCAUGAAA 2501 863 UGGGCUUC G GGAAGUUC 2167 GAACUUCC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GAAGCCCA 2502 864 GGGCUUCG G GAAGUUCG 2168 CGAACUUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGAAGCCC 2503 865 GGCUUCGG G AAGUUCGU 2169 ACGAACUU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCGAAGCC 2504 868 UUCGGGAA G UUCGUGCA 1353 UGCACGAA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UUCCCGAA 2505 872 GGAAGUUC G UGCAGUAC 1354 GUACUGCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GAACUUCC 2506 874 AAGUUCGU G CAGUACUG 1355 CAGUACUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACGAACUU 2507 877 UUCGUGCA G UACUGCCC 1356 GGGCAGUA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGCACGAA 2508 882 GCAGUACU G CCCCGGCA 1357 UGCCGGGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGUACUGC 2509 887 ACUGCCCC G GCACCUGG 2170 CCAGGUGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGGGCAGU 2510 888 CUGCCCCG G CACCUGGU 1358 ACCAGGUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGGGGCAG 2511 894 CGGCACCU G GUGCUUUA 2171 UAAAGCAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGGUGCCG 2512 895 GGCACCUG G UGCUUUAU 1359 AUAAAGCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAGGUGCC 2513 897 CACCUGGU G CUUUAUCC 1360 GGAUAAAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACCAGGUG 2514 907 UUUAUCCA G AUGGUCCA 2172 UGGACCAU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGGAUAAA 2515 910 AUCCAGAU G GUCCACGA 2173 UCGUGGAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AUCUGGAU 2516 911 UCCAGAUG G UCCACGAG 1361 CUCGUGGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAUCUGGA 2517 917 UGGUCCAC G AGGAGGGC 2174 GCCCUCCU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GUGGACCA 2518 919 GUCCACGA G GAGGGCUC 2175 GAGCCCUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCGUGGAC 2519 920 UCCACGAG G AGGGCUCG 2176 CGAGCCCU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CUCGUGGA 2520 922 CACGAGGA G GGCUCGCU 2177 AGCGAGCC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCCUCGUG 2521 923 ACGAGGAG G GCUCGCUG 2178 CAGCGAGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CUCCUCGU 2522 924 CGAGGAGG G CUCGCUGU 1362 ACAGCGAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCUCCUCG 2523 928 GAGGGCUC G CUGUCGGU 1363 ACCGACAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GAGCCCUC 2524 931 GGCUCGCU G UCGGUGCU 1364 AGCACCGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGCGAGCC 2525 934 UCGCUGUC G GUGCUGGG 2179 CCCAGCAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GACAGCGA 2526 935 CGCUGUCG G UGCUGGGG 1365 CCCCAGCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGACAGCG 2527 937 CUGUCGGU G CUGGGGUA 1366 UACCCCAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACCGACAG 2528 940 UCGGUGCU G GGGUACUC 2180 GAGUACCC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGCACCGA 2529 941 CGGUGCUG G GGUACUCU 2181 AGAGUACC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAGCACCG 2530 942 GGUGCUGG G GUACUCUG 2182 CAGAGUAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCAGCACC 2531 943 GUGCUGGG G UACUCUGU 1367 ACAGAGUA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CCCAGCAC 2532 950 GGUACUCU G UGCUCUAC 1368 GUAGAGCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGAGUACC 2533 952 UACUCUGU G CUCUACUC 1369 GAGUAGAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACAGAGUA 2534 963 CUACUCCA G CCUCAUGG 1370 CCAUGAGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGGAGUAG 2535 970 AGCCUCAU G GCGCUGCU 2183 AGCAGCGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AUGAGGCU 2536 971 GCCUCAUG G CGCUGCUG 1371 CAGCAGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAUGAGGC 2537 973 CUCAUGGC G CUGCUGGU 1372 ACCAGCAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCCAUGAG 2538 976 AUGGCGCU G CUGGUCCU 1373 AGGACCAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGCGCCAU 2539 979 GCGCUGCU G GUCCUCGC 2184 GCGAGGAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGCAGCGC 2540 980 CGCUGCUG G UCCUCGCC 1374 GGCGAGGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAGCAGCG 2541 986 UGGUCCUC G CCACCGUG 1375 CACGGUGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GAGGACCA 2542 992 UCGCCACC G UGCUGUGC 1376 GCACAGCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGUGGCGA 2543 994 GCCACCGU U CUGUGCAA 1377 UUGCACAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACGGUGGC 2544 997 ACCGUGCU G UGCAACCU 1378 AGGUUGCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGCACGGU 2545 999 CGUGCUGU G CAACCUCG 1379 CGAGGUUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACAGCACG 2546 1007 GCAACCUC U GCGCCAUG 2185 CAUGGCGC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GAGGUUGC 2547 1008 CAACCUCG G CGCCAUGC 1380 GCAUGGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGAGGUUG 2548 1010 ACCUCGGC G CCAUGCGC 1381 GCGCAUGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCCGAGGU 2549 1015 GGCGCCAU G CGCAACCU 1382 AGGUUUCU GGAGGAAACUCC CU UCAAUGACAUCGUCCGGG AUGGCGCC 2550 1017 CGCCAUUC G CAACCUCU 1383 AUAGGUUG GGAGGAAACUCC CU UCAAGGACAUCUUCCGGG GCAUGGCG 2551 1028 ACCUCUAU G CGAUGCAC 1384 GUGCAUCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AUAGAGGU 2552 1030 CUCUAUGC U AUGCACCG 2186 CGGUGCAU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCAUAGAG 2553 1033 UAUGCGAU G CACCGGCG 1385 CGCCGGUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AUCGCAUA 2554 1038 UAUGCACC G GCUGCUGC 2187 UCAGCCGC GUAGGAAACUCC CU UCAAGGACAUCGUCCGUG GGUGCAUC 2555 1039 AUGCACCG G CGUCUGCA 1386 UGCAGCCG GGAGUAAACUCC CU UCAAGGACAUCGUCCGUG CGGUGCAU 2556 1041 GCACCGGC U UCUGCAUC 2188 GCUGCAGC GGAGGAAACUCC CU UCAAUGACAUCGUCCGGG GCCGGUUC 2557 1042 CACCGUCG U CUUCAUCG 1387 CGCUGCAU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGCCGGUG 2558 1045 CGGCGUCU G CAUCUGCA 1388 UUCCGCUG GGAGGAAACUCC CU UCAAGUACAUCGUCCGGG AGCCGCCU 2559 1048 CUUCUGCA G CGGCACCC 1389 GGGUGCCG GGAGGAAACUCC CU UCAAGGACAUCGUCCUGG UGCAGCCG 2560 1050 GCUUCAGC U UCACCCGC 2189 GCGUGUGC UUAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCUGCAGC 2561 1051 CUGCAGCG G CACCCGCU 1390 CGCUGGUU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGU CGCUGCAG 2562 1057 CGUCACCC U CGCUCCUG 1391 CAGGAGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGGUGCCG 2563 1059 GCACCCUC U CUCCUGCA 1392 UGCAGGAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCGGGUGC 2564 1065 GCGCUCCU G CACCAGGG 1393 CCCUUGUG GGAUGAAACUCC CU UCAAGGACAUCGUCCGGG AGGAGCGC 2565 1071 CUGCACCA G GUACUGUG 2190 CACAGUCC UGAGUAAACUCC CU UCAAGUACAUCGUCCGGG UGGUGCAG 2566 1072 UGCACCAG U GACUGUGC 2191 GCACAUUC GGAGUAAACUCC CU UCAAGUACAUCGUCCGGG CUGGUGCA 2567 1073 GCACCAGG U ACUGUGCC 2192 GUCACAGU GGAGGAAACUCC CU UCAAGUACAUCGUCCGGG CCUGGUGC 2568 1077 CAGGGACU U UGCCGAGC 1394 GCUCGGCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGUCCCUG 2569 1079 GGGACUGU U CCGAGCCG 1395 CUGCUCGG UGAGGAAACUCC CU UCAAGGACAUCGUCCGGU ACAGUCCC 2570 1082 ACUUUUCC U AUCCUCUC 2193 GCUCGUCU UUAUGAAACUCC CU UCAAUGACAUCUUCCUGU UUCACAGU 2571 1084 UUUUCCUA U CCUCUCGC 1396 UCUCUCUG UUAUGAAACUCC CU UCAAUGACAUCUUCCUUU UCGUCACA 2572 1087 GCCGAGCC G CGCGCGGA 1397 UCCGCGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGCUCGGC 2573 1089 CGAGCCGC G CGCGGACG 1398 CGUCCGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCGGCUCG 2574 1091 AGCCGCGC G CGGACGGG 1399 CCCGUCCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCGCGGCU 2575 1093 CCGCGCGC G GACGGGAG 2194 CUCCCGUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCGCGCGG 2576 1094 CGCGCGCG G ACGGGAGG 2195 CCUCCCGU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGCGCGCG 2577 1097 GCGCGGAC G GGAGGGAA 2196 UUCCCUCC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GUCCGCGC 2578 1098 CGCGGACG C GAGGGAAG 2197 CUUCCCUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGUCCGCG 2579 1099 GCGGACGG C AGGGAAGC 2198 CCUUCCCU GCACGAAACUCC CU UCAAGCACAUCCUCCGGG CCCUCCGC 2580 1101 CGACCCGA C CCAAGCCU 2199 ACCCUUCC GCAGGAAACUCC CU UCAAGGACAUCGUCCCCG UCCCCUCC 2581 1102 CACGGCAC G GAAGCGUC 2200 GACGCUUC GGAGGAAACUCC CU UCAAGGACAUCGUCCCCG CUCCCGUC 2582 1103 ACCGCAGC C AACCGUCC 2201 GGACCCUU GCACGAAACUCC CU UCAAGCACAUCCUCCCCG CCUCCCGU 2583 1106 GGAGGCAA G CCUCCCCU 1400 AGCGCACG GGAGGAAACUCC CU UCAAGGACAUCGUCCCGG UUCCCUCC 2584 1108 AGGCAACC C UCCCCUCA 1401 UGAGGGCA GGAGGAAACUCC CU UCAACCACAUCGUCCCGG GCUUCCCU 2585 1117 UCCCCUCA C CCCCUGGA 1402 UCCACGGG GGACGAAACUCC CU UCAAGGACAUCGUCCCGG UCACCGGA 2586 1123 CAGCCCCU C CACGAGCU 2202 AGCUCCUC GGACGAAACUCC CU UCAAGGACAUCGUCCGGG ACGGGCUG 2587 1124 ACCCCCUG C ACGAGCUG 2203 CAGCUCCU GGAGGAAACUCC CU UCAAGGACAUCGUCCCCG CAGGCGCU 2588 1126 CCCCUCGA C GAGCUGGA 2204 UCCACCUC GCACGAAACUCC CU UCAAGGACAUCGUCCGCG UCCACGCG 2589 1127 CCCUCCAC C AGCUGGAU 2205 AUCCAGCU GCAGGAAACUCC CU UCAACGACAUCCUCCGCG CUCCAGGG 2590 1129 CUGGAGGA C CUGGAUCA 1403 UCAUCCAG GCACGAAACUCC CU UCAAGCACAUCGUCCGCG UCCUCCAG 2591 1132 GAGGAGCU C CAUCACCU 2206 ACCUGAUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACCUCCUC 2592 1133 AGCACCUC C AUCACCUC 2207 CACGUGAU GCACGAAACUCC CU UCAAGCACAUCCUCCGGG CACCUCCU 2593 1144 CACCUCCU C CUCCUGCC 1404 CCCACCAG GCACGAAACUCC CU UCAAGCACAUCCUCCGGG ACCAGGUG 2594 1147 CUCCUGCU C CUGGCGCU 1405 AGCGCCAG GGACGAAACUCC CU UCAAGGACAUCCUCCGGG ACCACGAG 2595 1150 CUCCUCCU C GCGCUGAU 2208 AUCACCCC GCAGGAAACUCC CU UCAAGCACAUCCUCCGCG ACCACCAG 2596 1151 UCCUCCUC C CGCUCAUG 1406 CAUCAGCG GCACGAAACUCC CU UCAAGGACAUCGUCCGGG CACCAGCA 2597 1153 CUCCUGGC C CUGAUGAC 1407 GUCAUCAG GCAGGAAACUCC CU UCAAGCACAUCCUCCGCG GCCAGCAG 2598 1156 CUCCCCCU C AUCACCCU 2209 ACCGUCAU GCAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACCGCCAG 2599 1159 CCCCUCAU C ACCCUCCU 2210 AGCACGCU GCACCAAACUCC CU UCAAGCACAUCGUCCGCG AUCACCCC 2600 1163 UGAUCACC C UGCUCUUC 1408 CAAGAGCA GCAGGAAACUCC CU UCAAGCACAUCCUCCGGG GCUCAUCA 2601 1165 AUGACCGU C CUCCUCAC 1409 GUCAAGAG GGAGCAAACUCC CU UCAAGCACAUCCUCCGCG ACGGUCAU 2602 1177 UUCACUAU C UGUUCUCU 1410 AGACAACA CGAGCAAACUCC CU UCAAGCACAUCCUCCGGG AUACUCAA 2603 1179 CACUAUGU C UUCUCUGC 1411 GCAGAGAA GGAGGAAACUCC CU UCAAGCACAUCCUCCGCG ACAUACUG 2604 1186 UGUUCUCU C CCCGUAAU 1412 AUUACGGC CGAGGAAACUCC CU UCAAGCACAUCCUCCCCG AGAGAACA 2605 1190 CUCUGCCC G UAAUUUAU 1413 AUAAAUUA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGGCAGAG 2606 1200 AAUUUAUC G CGCUUACU 1414 AGUAAGCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GAUAAAUU 2607 1202 UUUAUCGC G CUUACUAU 1415 AUAGUAAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCGAUAAA 2608 1211 CUUACUAU G CAGCAUUU 2211 AAAUGCUC GGAGGAAACUCC CU UCAAOGACAUCGUCCGGG AUAGUAAG 2609 1212 UUACUAUG G AGCAUUUA 2212 UAAAUGCU GGAGGAAACUCC CU UCAA0GACAUCGUCCG0G CAUAGUAA 2610 1214 ACUAUGGA G CAUUUAAG 1416 CUUAAAUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCCAUAGU 2611 1222 GCAUUUAA G GAUGUCAA 2213 UUGACAUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UUAAAUGC 2612 1223 CAUUUAAG G AUGUCAAG 2214 CUUGACAU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CUUAAAUG 2613 1226 UUAAGGAU G UCAAGGAG 1417 CUCCUUGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AUCCUUAA 2614 1231 GAUGUCAA G GAGAAAAA 2215 UUUUUCUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UUGACAUC 2615 1232 AUGUCAAG G AGAAAAAC 2216 GUUUUUCU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CUUGACAU 2616 1234 GUCAAGGA G AAAAACAG 2217 CUGUUUUU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCCUUGAC 2617 1242 GAAAAACA G GACCUCUG 2218 CAGAGGUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGUUUUUC 2618 1243 AAAAACAG G ACCUCUGA 2219 UCAGAGGU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CUGUUUUU 2619 1250 GGACCUCU G AAGAAGCA 2220 UGCUUCUU GGAGGAAACUCC CU UCAAGGACAUCGUCC0GG AGAGGUCC 2620 1253 CCUCUGAA G AAGCAGAA 2221 UUCUGCUU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UUCAGAGG 2621 1256 CUGAAGAA G CAGAAGAC 1418 GUCGUCUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UUCUUCAG 2622 1259 AAGAAGCA G AAGACCUC 2222 GAGGUCUU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGCUUCUU 2623 1262 AAGCAGAA G ACCUCCGA 2223 UCGGAGGU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UUCUGCUU 2624 1269 AGACCUCC G AGCCUUGC 2224 GCAAGGCU GGAGGAAACUCC CU UCAAG0ACAUCGUCCGGG GGAGGUCU 2625 1271 ACCUCCGA G CCUUGCGA 1419 UCGCAAGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCGGAGGU 2626 1276 CGAGCCUU G CGAUUUCU 1420 AGAAAUCG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AAGGCUCG 2627 1278 AGCCUUGC G AUUUCUAU 2225 AUAGAAAU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GCAAGGCU 2628 1289 UUCUAUCU G UGAUUUCA 1421 UGAAAUCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGAUAGAA 2629 1291 CUAUCUGU G AUUUCAAU 2226 AUUGAAAU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACAGAUAG 2630 1301 UUUCAAUU G UGGACCCU 1422 AGGGUCCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AAUUGAAA 2631 1303 UCAAUUGU G GACCCUUG 2227 CAAGGGUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACAAUUGA 2632 1304 CAAUUGUG G ACCCUUGG 2228 CCAAGGGU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CACAAUUG 2633 1311 GGACCCUU G GAUUUUUA 2229 UAAAAAUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AAGGGUCC 2634 1312 GACCCUUG G AUUUUUAU 2230 AUAAAAAU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAAGGGUC 2635 1329 CAUUUUCA G AUCUCCAG 2231 CUGGAGAU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGAAAAUG 2636 1337 GAUCUCCA G UAUUUCGG 1423 CCGAAAUA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGGAGAUC 2637 1344 AGUAUUUC G GAUAUUUU 2232 AAAAUAUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GAAAUACU 2638 1345 GUAUUUCG G AUAUUUUU 2233 AAAAAUAU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGAAAUAC 2639 1360 UUUCACAA G AUUUUCAU 2234 AUGAAAAU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UUGUGAAA 2640 1371 UUUCAUUA G ACCUCUUA 2235 UAAGAGGU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UAAUGAAA 2641 1380 ACCUCUUA G GUACAGGA 2236 UCCUGUAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UAAGAGGU 2642 1381 CCUCUUAG G UACAGGAG 1424 CUCCUGUA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CUAAGAGG 2643 1386 UAGGUACA G GAGCCGGU 2237 ACCGGCUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGUACCUA 2644 1387 AGGUACAG G AGCCGGUG 2238 CACCGGCU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CUGUACCU 2645 1389 GUACAGGA G CCGGUGCA 1425 UGCACCGG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCCUGUAC 2646 1392 CAGGAGCC G GUGCAGCA 2239 UGCUGCAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG GGCUCCUG 2647 1393 AGGAGCCG G UGCAGCAA 1426 UUGCUGCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CGGCUCCU 2648 1395 GAGCCGGU G CAGCAAUU 1427 AAUUGCUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACCGGCUC 2649 1398 CCGGUGCA G CAAUUCCA 1428 UGGAAUUG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGCACCGG 2650 1414 ACUAACAU G GAAUCCAG 2240 CUGGAUUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AUGUUAGU 2651 1415 CUAACAUG G AAUCCAGU 2241 ACUGGAUU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CAUGUUAG 2652 1422 GGAAUCCA G UCUGUGAC 1429 GUCACAGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGGAUUCC 2653 1426 UCCAGUCU G UGACAGUG 1430 CACUGUCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGACUGGA 2654 1428 CAGUCUGU G ACAGUGUU 2242 AACACUGU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACAGACUG 2655 1432 CUGUGACA G UGUUUUUC 1431 GAAAAACA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGUCACAG 2656 1434 GUGACAGU G UUUUUCAC 1432 GUGAAAAA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACUGUCAC 2657 1446 UUCACUCU G UGGUAAGC 1433 GCUUACCA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGAGUGAA 2658 1448 CACUCUGU G GUAAGCUG 2243 CAGCUUAC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG ACAGAGUG 2659 1449 ACUCUGUG G UAAGCUGA 1434 UCAGCUUA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CACAGAGU 2660 1453 UGUGGUAA G CUGAGGAA 1435 UUCCUCAG GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UUACCACA 2661 1456 GGUAAGCU G AGGAAUAU 2244 AUAUUCCU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AGCUUACC 2662 1458 UAAGCUGA G GAAUAUGU 2245 ACAUAUUC GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UCAGCUUA 2663 1459 AAGCUGAG G AAUAUGUC 2246 GACAUAUU GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG CUCAGCUU 2664 1465 AGGAAUAU G UCACAUUU 1436 AAAUGUGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG AUAUUCCU 2665 1477 CAUUUUCA G UCAAAGAA 1437 UUCUUUGA GGAGGAAACUCC CU UCAAGGACAUCGUCCGGG UGAAAAUG 2666 

What we claim is:
 1. A nucleic acid molecule that down regulates expression of a prostaglandin D2 receptor (PTGDR) gene.
 2. The nucleic acid molecule of claim 1, wherein said nucleic acid molecule is an enzymatic nucleic acid molecule.
 3. The nucleic acid molecule of claim 1, wherein said nucleic acid molecule is an antisense nucleic acid molecule.
 4. The enzymatic nucleic acid molecule of claim 2, wherein said enzymatic nucleic acid molecule comprises a sequence selected from the group of sequences consisting of SEQ ID NOs: 228-454, 831-1206, 1438-1668, 1715-2057, and 2247-2666.
 5. The enzymatic nucleic acid molecule of claim 2, wherein said enzymatic nucleic acid molecule comprises at least one binding arm wherein one or more of said binding arms comprises a sequence complementary to a sequence selected from the group of sequences consisting of SEQ ID NOs: 1-227, 455-830, 1207-1437, 1669-1714, and 2058-2246.
 6. The antisense nucleic acid molecule of claim 3, wherein said antisense nucleic acid molecule comprises a sequence complementary to a sequence selected from the group of sequences consisting of SEQ ID NOs: 1-227, 455-830, 1207-1437, 1669-1714, and 2058-2246.
 7. The nucleic acid molecule of claim 1, wherein said nucleic acid molecule is adapted to treat asthma.
 8. The nucleic acid molecule of claim 1, wherein said nucleic acid molecule comprises at least one 2′-sugar modification.
 9. The nucleic acid molecule of claim 1, wherein said nucleic acid molecule comprises at least one phosphate backbone modification.
 10. A method of reducing PTGDR activity in a cell, comprising contacting said cell with the nucleic acid molecule of claim 1 under conditions suitable for said reduction.
 11. A method of treatment of a patient having a condition associated with the level of PTGDR, comprising contacting cells of said patient with the nucleic acid molecule of claim 1, under conditions suitable for said treatment.
 12. The method of claim 11 further comprising the use of one or more drug therapies under conditions suitable for said treatment.
 13. A pharmaceutical composition comprising an enzymatic nucleic acid molecule of claim
 1. 14. A method of administering to a mammal the nucleic acid molecule of claim 1, comprising contacting said mammal with the molecule under conditions suitable for said administration.
 15. The method of claim 14, wherein said mammal is a human.
 16. The method of claim 14 wherein said administration is in the presence of a delivery reagent.
 17. The method of claim 16, wherein said delivery reagent is a lipid.
 18. The method of claim 17, wherein said lipid is a cationic lipid.
 19. The method of claim 17, wherein said lipid is a phospholipid.
 20. The method of claim 17, wherein said delivery reagent is a liposome. 